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Sample records for aspergillus niger growing

  1. ASPERGILLUS NIGER ASPERGILLUS NIGER

    African Journals Online (AJOL)

    eobe

    Additives such as low molecular weight alcohols, trace metals, phytate, lipids etc have been reported to stimulate citric acid production. Hence the objective of this study was to investigate the effect of stimulating the metabolic activity of activity of Aspergillus niger for the purpose of improved citric acid production from ...

  2. The intra- and extracellular proteome of Aspergillus niger growing on defined medium with xylose or maltose as carbon substrate

    Science.gov (United States)

    2010-01-01

    Background The filamentous fungus Aspergillus niger is well-known as a producer of primary metabolites and extracellular proteins. For example, glucoamylase is the most efficiently secreted protein of Aspergillus niger, thus the homologous glucoamylase (glaA) promoter as well as the glaA signal sequence are widely used for heterologous protein production. Xylose is known to strongly repress glaA expression while maltose is a potent inducer of glaA promoter controlled genes. For a more profound understanding of A. niger physiology, a comprehensive analysis of the intra- and extracellular proteome of Aspergillus niger AB1.13 growing on defined medium with xylose or maltose as carbon substrate was carried out using 2-D gel electrophoresis/Maldi-ToF and nano-HPLC MS/MS. Results The intracellular proteome of A. niger growing either on xylose or maltose in well-aerated controlled bioreactor cultures revealed striking similarities. In both cultures the most abundant intracellular protein was the TCA cycle enzyme malate-dehydrogenase. Moreover, the glycolytic enzymes fructose-bis-phosphate aldolase and glyceraldehyde-3-phosphate-dehydrogenase and the flavohemoglobin FhbA were identified as major proteins in both cultures. On the other hand, enzymes involved in the removal of reactive oxygen species, such as superoxide dismutase and peroxiredoxin, were present at elevated levels in the culture growing on maltose but only in minor amounts in the xylose culture. The composition of the extracellular proteome differed considerably depending on the carbon substrate. In the secretome of the xylose-grown culture, a variety of plant cell wall degrading enzymes were identified, mostly under the control of the xylanolytic transcriptional activator XlnR, with xylanase B and ferulic acid esterase as the most abundant ones. The secretome of the maltose-grown culture did not contain xylanolytic enzymes, instead high levels of catalases were found and glucoamylase (multiple spots) was

  3. The intra- and extracellular proteome of Aspergillus niger growing on defined medium with xylose or maltose as carbon substrate

    Directory of Open Access Journals (Sweden)

    Wissing Josef

    2010-04-01

    Full Text Available Abstract Background The filamentous fungus Aspergillus niger is well-known as a producer of primary metabolites and extracellular proteins. For example, glucoamylase is the most efficiently secreted protein of Aspergillus niger, thus the homologous glucoamylase (glaA promoter as well as the glaA signal sequence are widely used for heterologous protein production. Xylose is known to strongly repress glaA expression while maltose is a potent inducer of glaA promoter controlled genes. For a more profound understanding of A. niger physiology, a comprehensive analysis of the intra- and extracellular proteome of Aspergillus niger AB1.13 growing on defined medium with xylose or maltose as carbon substrate was carried out using 2-D gel electrophoresis/Maldi-ToF and nano-HPLC MS/MS. Results The intracellular proteome of A. niger growing either on xylose or maltose in well-aerated controlled bioreactor cultures revealed striking similarities. In both cultures the most abundant intracellular protein was the TCA cycle enzyme malate-dehydrogenase. Moreover, the glycolytic enzymes fructose-bis-phosphate aldolase and glyceraldehyde-3-phosphate-dehydrogenase and the flavohemoglobin FhbA were identified as major proteins in both cultures. On the other hand, enzymes involved in the removal of reactive oxygen species, such as superoxide dismutase and peroxiredoxin, were present at elevated levels in the culture growing on maltose but only in minor amounts in the xylose culture. The composition of the extracellular proteome differed considerably depending on the carbon substrate. In the secretome of the xylose-grown culture, a variety of plant cell wall degrading enzymes were identified, mostly under the control of the xylanolytic transcriptional activator XlnR, with xylanase B and ferulic acid esterase as the most abundant ones. The secretome of the maltose-grown culture did not contain xylanolytic enzymes, instead high levels of catalases were found and

  4. Effects of Aspergillus niger fermented rapeseed meal on nutrient digestibility, growth performance and serum parameters in growing pigs.

    Science.gov (United States)

    Shi, Changyou; He, Jun; Wang, Jianping; Yu, Jie; Yu, Bing; Mao, Xiangbing; Zheng, Ping; Huang, Zhiqing; Chen, Daiwen

    2016-04-01

    The aim of the present study was to investigate the influences of Aspergillus niger fermented rapeseed meal (FRSM) on growth performance and nutrient digestibility of growing pigs. A total of 72 growing pigs (body weight = 40.8 ± 2.1 kg) were used in feeding trials, lasting for up to 42 days, and were randomly allotted to one of three diets, including a corn-soybean meal control diet as well as two experimental diets containing 10% unfermented rapeseed meal (RSM) or 10% FRSM. The results showed that average daily gain and feed conversion ratio of pigs fed FRSM were superior (P digestibility for dry matter, protein, calcium and phosphorus than pigs fed unfermented RSM diet and did not differ from the FRSM diet. Pigs fed FRSM had lower levels (P digestibility of RSM for pigs and FRSM is a promising alternative protein for pig production. © 2015 Japanese Society of Animal Science.

  5. The potential of Aspergillus fumigatus and Aspergillus niger in ...

    African Journals Online (AJOL)

    Fungi isolated included Aspergillus candidus, A. fumigatus, A. niger, A. tamarii, Mucor rouxii, Penicillium notatum and Rhizopus sp. Aspergillus fumigatus and Aspergillus niger were selected for heavy metals bioaccumulation studies on PDB-amended with lagoon water in ratios of 1:1, 1:3 and 1:5 respectively for 3 weeks.

  6. Shedding light on Aspergillus niger volatile exometabolome

    National Research Council Canada - National Science Library

    Costa, Carina Pedrosa; Gonçalves Silva, Diogo; Rudnitskaya, Alisa; Almeida, Adelaide; Rocha, Sílvia M

    2016-01-01

    An in-depth exploration of the headspace content of Aspergillus niger cultures was performed upon different growth conditions, using a methodology based on advanced multidimensional gas chromatography...

  7. Aspergillus niger contains the cryptic phylogenetic species A. awamori

    DEFF Research Database (Denmark)

    Perrone, Giancarlo; Stea, Gaetano; Epifani, Filomena

    2011-01-01

    Aspergillus section Nigri is an important group of species for food and medical mycology, and biotechnology. The Aspergillus niger ‘aggregate’ represents its most complicated taxonomic subgroup containing eight morphologically indistinguishable taxa: A. niger, Aspergillus tubingensis, Aspergillus...

  8. Sporulation inhibited secretion in Aspergillus niger

    NARCIS (Netherlands)

    Krijgsheld, P.

    2013-01-01

    Aspergillus niger is abundantly found in nature. It degrades dead material of plants and animals but can also be a pathogen of these organisms. Aspergillus niger is also important for mankind because it is one of the main organisms used for the industrial production of enzymes. These enzymes are

  9. Morphogenesis and protein production in Aspergillus niger

    NARCIS (Netherlands)

    Kwon, Min Jin

    2014-01-01

    The research described in this thesis aims to get more fundamental insights in the molecular mechanisms used by Aspergillus niger in relation to control morphology and protein secretion. Knowledge on these two aspects is highly relevant to further optimization of A.niger as a cell factory

  10. Biotransformation of Stypotriol triacetate by Aspergillus niger

    Science.gov (United States)

    Areche, Carlos; Vaca, Inmaculada; Labbe, Pamela; Soto-Delgado, Jorge; Astudillo, Luis; Silva, Mario; Rovirosa, Juana; San-Martin, Aurelio

    2011-07-01

    Biological transformation of the meroditerpenoid, stypotriol triacetate ( 1) by the fungi Aspergillus niger, Cunninghamella elegans, Gibberella fujikuroi and Mucor plumbeus was studied. The incubation of 1 with A. niger yielded the new compound 6',14-diacetoxy-stypol-4,5-dione ( 2) whose structure was established by 1H, 13C and 2D NMR and supported by DFT/GIAO.

  11. GLUCOSIDASE GENE FROM ASPERGILLUS NIGER F321

    African Journals Online (AJOL)

    Richard Auta

    One of the cellulases produced in this fashion by species of Aspergillus is β- glucosidase. .... the crude enzyme (sample) or nutrient broth (Control) was placed and incubated for 2 .... 7: Multiple sequence alignment of β-glucosidases amino acids from Aspergillus niger AnBg11, ANRA12.6 and ANRA12.9. [Symbols: Catalytic ...

  12. Changes in dehydrogenases activity, number and ultrastructure of mitochondria in Aspergillus niger mycelium growing on molasses media.

    Science.gov (United States)

    Gabara, B; Zakowska, Z

    1993-01-01

    Effects of toxic molasses compounds and of the antifoamer (Spumol BJ) on dehydrogenases activity, on the number and ultrastructure of mitochondria in A. niger mycelium of two strains characterized by different tolerance to toxic agents, were observed. In spite of significantly higher dehydrogenases activity in the intolerant strain (R-16) mycelia developing both on productive molasses in the presence of the defoamer and on non-productive molasses are characterized by marked reduction in the activity of these enzymes. Changes in respiratory enzymes activity are partly correlated with the number of mitochondria but mostly with abnormalities in their ultrastructure.

  13. Characterisation of Aspergillus niger prolyl aminopeptidase

    NARCIS (Netherlands)

    Basten, E.J.W.; Moers, A.P.H.A.; Ooyen, van A.J.J.; Schaap, P.J.

    2005-01-01

    We have cloned a gene (papA) that encodes a prolyl aminopeptidase from Aspergillus niger. Homologous genes are present in the genomes of the Eurotiales A. nidulans, A. fumigatus and Talaromyces emersonii, but the gene is not present in the genome of the yeast Saccharomyces cerevisiae. Cell extracts

  14. Identification of thermostable beta-xylosidase activities produced by Aspergillus brasiliensis and Aspergillus niger

    DEFF Research Database (Denmark)

    Pedersen, Mads; Lauritzen, H.K.; Frisvad, Jens Christian

    2007-01-01

    Twenty Aspergillus strains were evaluated for production of extracellular cellulolytic and xylanolytic activities. Aspergillus brasiliensis, A. niger and A. japonicus produced the highest xylanase activities with the A. brasiliensis and A. niger strains producing thermostable beta...

  15. Identification of thermostable β-xylosidase activities produced by Aspergillus brasiliensis and Aspergillus niger

    DEFF Research Database (Denmark)

    Pedersen, Mads; Lauritzen, Henrik Klitgaard; Frisvad, Jens Christian

    2007-01-01

    Twenty Aspergillus strains were evaluated for production of extracellular cellulolytic and xylanolytic activities. Aspergillus brasiliensis, A. niger and A. japonicus produced the highest xylanase activities with the A. brasiliensis and A. niger strains producing thermostable beta...

  16. 21 CFR 173.120 - Carbohydrase and cellulase derived from Aspergillus niger.

    Science.gov (United States)

    2010-04-01

    ... cellulase derived from Aspergillus niger. Carbohydrase and cellulase enzyme preparation derived from Aspergillus niger may be safely used in food in accordance with the following prescribed conditions: (a) Aspergillus niger is classified as follows: Class, Deuteromycetes; order, Moniliales; family, Moniliaceae...

  17. [Induced synthesis of hydroxyapatite by Aspergillus niger].

    Science.gov (United States)

    He, Fei; Lian, Bin; Liu, Shirong; Gong, Guohong

    2011-03-01

    The research objective is to induce hydroxyapatite (HAP) synthesis by using fungus. We used the PDA (Potato Dextrose Agar Medium) liquid medium containing different concentrations of Na2HPO4 and CaCO3 to study the way Aspergillus niger synthesize HAP, to observe the induced mineral crystal structure and to analyze the induced mineral type with transmission electron microscopy (TEM) and X-ray diffraction (XRD). The main results are as follows: (1) A. niger can induce HAP synthesis in PDA liquid medium with the proper concentration of Na2HPO4 and CaCO3. (2) The reaction of A. niger inducing HAP synthesis depends on the time of the response system. Longer time is more advantageous in producing HAP. The main reason for A. niger inducing HAP crystals formation are as follows: fungal metabolism produces the acidic substances to dissolve CaCO, and the growth mycelia absorbing Ca2+ lead to Ca2+ enriched on the surface, to promote the production of secondary mineral apatite and further transform into HAP in the mycelia spheres. A. niger can inducing HAP crystals formation in PDA liquid medium containing Na2HPO4 and CaCO3. Considering the importance of HAP in bio-medicine materials and its costly prices, our method for HAP bio-induction is of the temperate response condition, simple preparation craft, and lower cost, which has the potential application prospect.

  18. Assessment of Aspergillus niger biofilm growth kinetics in ...

    African Journals Online (AJOL)

    Jane

    2011-10-12

    Oct 12, 2011 ... assessed by evaluating the CO2 released during the fermentation in minibioreactors. Key words: Aspergillus niger, biofilm, mathematical modeling, endogenous respiration, Cryo-SEM. INTRODUCTION. Aspergillus niger is currently one of the microbial species of main biotechnological importance because ...

  19. screening and improvement of local isolates of aspergillus niger for ...

    African Journals Online (AJOL)

    DR. AMINU

    ABSTRACT. The study involved the screening of fourteen isolates of Aspergillus niger for citric acid production from glucose. The study was aimed at screening and improving local strains of Aspergillus niger with potential for citric acid production. All the isolates screened produced varying amounts of citric acid, the highest ...

  20. Hydrolysis of raw tuber starches by amylase of Aspergillus niger ...

    African Journals Online (AJOL)

    Hydrolysis of raw tuber starches by amylase of Aspergillus niger AM07 isolated from the soil. ... The crude amylase preparation of A. niger AM07 had temperature and pH optima activities at 60oC and 4.0 respectively. The optimum substrate concentration was 3 %. The action of the crude amylase of A. niger on raw tuber ...

  1. Aspergillus niger contains the cryptic phylogenetic species A. awamori.

    Science.gov (United States)

    Perrone, Giancarlo; Stea, Gaetano; Epifani, Filomena; Varga, János; Frisvad, Jens C; Samson, Robert A

    2011-11-01

    Aspergillus section Nigri is an important group of species for food and medical mycology, and biotechnology. The Aspergillus niger 'aggregate' represents its most complicated taxonomic subgroup containing eight morphologically indistinguishable taxa: A. niger, Aspergillus tubingensis, Aspergillus acidus, Aspergillus brasiliensis, Aspergillus costaricaensis, Aspergillus lacticoffeatus, Aspergillus piperis, and Aspergillus vadensis. Aspergillus awamori, first described by Nakazawa, has been compared taxonomically with other black aspergilli and recently it has been treated as a synonym of A. niger. Phylogenetic analyses of sequences generated from portions of three genes coding for the proteins β-tubulin (benA), calmodulin (CaM), and the translation elongation factor-1 alpha (TEF-1α) of a population of A. niger strains isolated from grapes in Europe revealed the presence of a cryptic phylogenetic species within this population, A. awamori. Morphological, physiological, ecological and chemical data overlap occurred between A. niger and the cryptic A. awamori, however the splitting of these two species was also supported by AFLP analysis of the full genome. Isolates in both phylospecies can produce the mycotoxins ochratoxin A and fumonisin B₂, and they also share the production of pyranonigrin A, tensidol B, funalenone, malformins, and naphtho-γ-pyrones. In addition, sequence analysis of four putative A. awamori strains from Japan, used in the koji industrial fermentation, revealed that none of these strains belong to the A. awamori phylospecies. Copyright © 2011 British Mycological Society. Published by Elsevier Ltd. All rights reserved.

  2. Expression of the Aspergillus terreus itaconic acid biosynthesis cluster in Aspergillus niger

    NARCIS (Netherlands)

    Straat, van der L.; Vernooij, M.; Lammers, M.; Berg, van den W.A.M.; Schonewille, T.; Cordewener, J.; Meer, van der I.; Koops, A.J.; Graaff, de L.H.

    2014-01-01

    BACKGROUND: Aspergillus terreus is a natural producer of itaconic acid and is currently used to produce itaconic acid on an industrial scale. The metabolic process for itaconic acid biosynthesis is very similar to the production of citric acid in Aspergillus niger. However, a key enzyme in A. niger,

  3. Effect of simulated microgravity on Aspergillus niger

    Science.gov (United States)

    Pratap, Jeffrey J.

    2005-08-01

    A rotating bioreactor was developed to simulate microgravity and its influence was studied on fungal growth. The reactor was designed to simulate microgravity using 'free fall' principle, which creates an apparent weightlessness for a brief period of time. In this experiment, a sealed vertically rotating tube is the reactor in which the cells are grown. For the first time vertically rotating tubes were used to obtain 'free fall' thereby simulating microgravity. Simulated microgravity served significant in the alteration of growth and productivity of Aspergillus niger, a common soil fungi. Two other sets of similar cultures were maintained as still and shake control cultures to compare with the growth and productivity of cells in rotating culture. It was found increased growth and productivity occurred in simulated microgravity. Since this experiment involves growth of cells in a liquid medium, the fluidic effects must also be studied which is a limitation.

  4. Reconstruction of the central carbon metabolism of Aspergillus niger

    DEFF Research Database (Denmark)

    David, Helga; Åkesson, Mats Fredrik; Nielsen, Jens

    2003-01-01

    The topology of central carbon metabolism of Aspergillus niger was identified and the metabolic network reconstructed, by integrating genomic, biochemical and physiological information available for this microorganism and other related fungi. The reconstructed network may serve as a valuable...

  5. Pectinolytic complex production by Aspergillus niger URM 4645 ...

    African Journals Online (AJOL)

    -PG), pectin lyase (PL), and pectin methylesterase (PE), produced by Aspergillus niger URM 4645, were studied in solid state fermentation (SSF) using yellow passion fruit peels as substrate. The effect of substrate amount, initial moisture ...

  6. The het-c heterokaryon incompatibility gene in Aspergillus niger

    NARCIS (Netherlands)

    Diepeningen, van A.D.; Pal, K.; Lee, T.; Hoekstra, R.F.; Debets, A.J.M.

    2009-01-01

    Heterokaryon incompatibility among Aspergillus niger strains is a widespread phenomenon that is observed as the inability to form stable heterokaryons. The genetic basis of heterokaryon incompatibility reactions is well established in some sexual filamentous fungi but largely unknown in presumed

  7. On the safety of Aspergillus niger - a review

    DEFF Research Database (Denmark)

    Schuster, E.; Dunn-Coleman, N.; Frisvad, Jens Christian

    2002-01-01

    Administration. In addition, A. niger is used for biotransformations and waste treatment. In the last two decades, A. niger has been developed as an important transformation host to over-express food enzymes. Being pre-dated by older names, the name A. niger has been conserved for economical and information......Aspergillus niger is one of the most important microorganisms used in biotechnology. It has been in use already for many decades to produce extracellular (food) enzymes and citric acid. In fact, citric acid and many A. niger enzymes are considered GRAS by the United States Food and Drug...

  8. Production and Purification of Peroxidase from Aspergillus niger.

    OpenAIRE

    Mohammed A. Jebor

    2017-01-01

    This study was conducted in the laboratories of Biology Department, College of Science, which deals with isolation and purification of peroxidase and optimization of process parameters to achieve maximum yield of peroxidase by Aspergillus niger. Solid-state fermentation of Aspergillus niger was carried out for enhanced production of peroxidase using hydrogen peroxide as the substrate of enzyme maximum activity of the enzyme was achieved under optimum growth conditions. The optimum conditi...

  9. Partially purified polygalacturonase from Aspergillus niger (SA6 ...

    African Journals Online (AJOL)

    Polygalacturonase (PG) was isolated from Aspergillus niger (A. niger) (SA6), partially purified and characterized. The PG showed two bands on SDS-PAGE suggesting an “endo and exo PG with apparent molecular weights of 35 and 40 KDa, respectively. It was purified 9-fold with a yield of 0.18% and specific activity of 246 ...

  10. Organic acid production in Aspergillus niger and other filamentous fungi

    NARCIS (Netherlands)

    Odoni, Dorett I.

    2017-01-01

    The aim of the thesis was to increase the understanding of organic acid production in Aspergillus niger and other filamentous fungi, with the ultimate purpose to improve A. niger as biotechnological production host. In Chapter 1, the use of microbial cell-factories for the

  11. A novel pig feed formulation containing Aspergillus niger CSA35 ...

    African Journals Online (AJOL)

    This study investigated the effects of Aspergillus niger CSA35 pretreated-cassava (Manihot esculenta Crantz) peel feed (CPFG) on the body weight gain and some selected biochemical parameters of pigs. Cassava peels treated with biomass of A. niger CSA35 for a period of three weeks to initiate enzymatic digestion of ...

  12. Ecophysiological characterization of Aspergillus carbonarius, Aspergillus tubingensis and Aspergillus niger isolated from grapes in Spanish vineyards.

    Science.gov (United States)

    García-Cela, E; Crespo-Sempere, A; Ramos, A J; Sanchis, V; Marin, S

    2014-03-03

    The aim of this study was to evaluate the diversity of black aspergilli isolated from berries from different agroclimatic regions of Spain. Growth characterization (in terms of temperature and water activity requirements) of Aspergillus carbonarius, Aspergillus tubingensis and Aspergillus niger was carried out on synthetic grape medium. A. tubingensis and A. niger showed higher maximum temperatures for growth (>45 °C versus 40-42 °C), and lower minimum aw requirements (0.83 aw versus 0.87 aw) than A. carbonarius. No differences in growth boundaries due to their geographical origin were found within A. niger aggregate isolates. Conversely, A. carbonarius isolates from the hotter and drier region grew and produced OTA at lower aw than other isolates. However, little genetic diversity in A. carbonarius was observed for the microsatellites tested and the same sequence of β-tubulin gene was observed; therefore intraspecific variability did not correlate with the geographical origin of the isolates or with their ability to produce OTA. Climatic change prediction points to drier and hotter climatic scenarios where A. tubingensis and A. niger could be even more prevalent over A. carbonarius, since they are better adapted to extreme high temperature and drier conditions. Copyright © 2013 Elsevier B.V. All rights reserved.

  13. Switching from a unicellular to multicellular organization in an Aspergillus niger hypha

    NARCIS (Netherlands)

    Bleichrodt, Robert-Jan; Hulsman, Marc; Wosten, H.A.B.; Reinders, Marcel J T

    UNLABELLED: Pores in fungal septa enable cytoplasmic streaming between hyphae and their compartments. Consequently, the mycelium can be considered unicellular. However, we show here that Woronin bodies close ~50% of the three most apical septa of growing hyphae of Aspergillus niger. The incidence of

  14. Effects of Aspergillus niger treated Shea butter cake based diets on ...

    African Journals Online (AJOL)

    Effects of feed intake, weight gain and digestibility when growing Red Sokoto goats consuming Aspergillus niger treated and untreated shea-butter cake (SBC) were determined. Twenty five Red Sokoto goats in a completely randomized design model with 56 d periods consumed diet A (control, without SBC), B (15% ...

  15. Use of Aspergillus niger for bioconversion of apple distillery waste

    Energy Technology Data Exchange (ETDEWEB)

    Friedrich, J.; Cimerman, A.; Perdih, A.

    1983-01-01

    The bioconversion of waste material remaining after apple brandy distillation was investigated. Different cellulolytic fungi were tested for their ability to convert the waste organic substances into microbial biomass. An Aspergillus niger strain was chosen as the most convenient microorganism. By growing this mold on the apple slop the following results were obtained: filtration time was shortened by 30 times, reduction of the chemical oxygen demand in the liquid phase in the range of 50-80% depending on the substrate dilution and a dry filter cake enriched with fungal biomass to about 12 g/l containing up to 22% raw proteins and certain amounts of cellulolytic enzymes in the filtrate. The influence of the initial pH, the salt addition and the dilution of the substrate were studied as well. 12 references.

  16. Aspergillus Niger Genomics: Past, Present and into the Future

    Energy Technology Data Exchange (ETDEWEB)

    Baker, Scott E.

    2006-09-01

    Aspergillus niger is a filamentous ascomycete fungus that is ubiquitous in the environment and has been implicated in opportunistic infections of humans. In addition to its role as an opportunistic human pathogen, A. niger is economically important as a fermentation organism used for the production of citric acid. Industrial citric acid production by A. niger represents one of the most efficient, highest yield bioprocesses in use currently by industry. The genome size of A. niger is estimated to be between 35.5 and 38.5 megabases (Mb) divided among eight chromosomes/linkage groups that vary in size from 3.5 - 6.6 Mb. Currently, there are three independent A. niger genome projects, an indication of the economic importance of this organism. The rich amount of data resulting from these multiple A. niger genome sequences will be used for basic and applied research programs applicable to fermentation process development, morphology and pathogenicity.

  17. [Hypersensitivity pneumonitis induced by Aspergillus niger--a case report].

    Science.gov (United States)

    Miyazaki, Hiroo; Gemma, Hitoshi; Uemura, Keiichi; Ono, Takahisa; Masuda, Masafumi; Sano, Takehisa; Sato, Masaki; Koshimizu, Naoki; Suda, Takafumi; Chida, Kingo

    2004-07-01

    A 52-year-old woman was hospitalized because of severe cough in August 1994. She had engaged in culturing roses in greenhouses since 1968, and had developed a cough during the summer of 1990. Chest radiography showed diffuse ground-glass opacity in both lung fields, and she suffered from hypoxemia (PaO2 = 45.6 torr) while breathing room air. The lymphocyte count in the bronchoalveolar lavage fluid was increased, and transbronchial lung biopsy specimens showed lymphocyte alveolitis in the alveolar spaces. After admission, the patient's symptoms improved rapidly without medication. However, on her return to work, the cough and hypoxemia reappeared. In her rose culture, she had used Rockwool, and Aspergillus niger was detected predominantly in the Rockwool. Precipitins against the extracts of Aspergillus niger were detected with the double immunodiffusion test and the inhalation provocation test yielded clinical symptoms. Our diagnosis was hypersensitivity pneumonitis caused by Aspergillus niger.

  18. Electrochemical monitoring of citric acid production by Aspergillus niger

    Energy Technology Data Exchange (ETDEWEB)

    Kutyła-Olesiuk, Anna; Wawrzyniak, Urszula E.; Ciosek, Patrycja; Wróblewski, Wojciech, E-mail: wuwu@ch.pw.edu.pl

    2014-05-01

    Highlights: • Citric acid fermentation process (production) by Aspergillus niger. • Qualitative/quantitative monitoring of standard culture and culture infected with yeast. • Electronic tongue based on potentiometric and voltammetric sensors. • Evaluation of the progress and the correctness of the fermentation process. • The highest classification abilities of the hybrid electronic tongue. - Abstract: Hybrid electronic tongue was developed for the monitoring of citric acid production by Aspergillus niger. The system based on various potentiometric/voltammetric sensors and appropriate chemometric techniques provided correct qualitative and quantitative classification of the samples collected during standard Aspergillus niger culture and culture infected with yeast. The performance of the proposed approach was compared with the monitoring of the fermentation process carried out using classical methods. The results obtained proved, that the designed hybrid electronic tongue was able to evaluate the progress and correctness of the fermentation process.

  19. Phosphate solubilizing ability of two Arctic Aspergillus niger strains

    Directory of Open Access Journals (Sweden)

    Shiv Mohan Singh,

    2011-06-01

    Full Text Available Many filamentous fungi were isolated from the soils of Ny-Ålesund, Spitsbergen, Svalbard, and were screened in vitro for their phosphate solubilizing ability. Two strains of Aspergillus niger showed good tricalcium phosphate (TCP solubilizing ability in Pikovskaya's medium. The TCP solubilization index was calculated at varying levels of pH and temperatures. The ability of Aspergillus niger strain-1 to solubilize and release inorganic-P was 285 µg ml–1, while Aspergillus niger strain-2 solubilized 262 µg ml–1 from 0.5% TCP after seven days. This is the first report of TCP solubilization by Arctic strains that may serve as very good phosphate solubilizers in the form of biofertilizer.

  20. Aconitase and citric acid fermentation by Aspergillus niger.

    OpenAIRE

    Kubicek, C P; Röhr, M

    1985-01-01

    In view of the often-cited theory that citric acid accumulation is caused by an inhibition of aconitase activity, the equilibrium of the reaction of aconitase was investigated by comparing in vivo steady-state concentrations of citrate and isocitrate in Aspergillus niger grown under various citric acid-producing conditions. With the equilibrium catalyzed by the A. niger enzyme in vitro, similar values were obtained. The validity of our in vivo measurements was verified by the addition of the ...

  1. Electrochemical monitoring of citric acid production by Aspergillus niger.

    Science.gov (United States)

    Kutyła-Olesiuk, Anna; Wawrzyniak, Urszula E; Ciosek, Patrycja; Wróblewski, Wojciech

    2014-05-01

    Hybrid electronic tongue was developed for the monitoring of citric acid production by Aspergillus niger. The system based on various potentiometric/voltammetric sensors and appropriate chemometric techniques provided correct qualitative and quantitative classification of the samples collected during standard Aspergillus niger culture and culture infected with yeast. The performance of the proposed approach was compared with the monitoring of the fermentation process carried out using classical methods. The results obtained proved, that the designed hybrid electronic tongue was able to evaluate the progress and correctness of the fermentation process. Copyright © 2014 Elsevier B.V. All rights reserved.

  2. Fumonisins in Aspergillus niger: Industrial and food aspects

    DEFF Research Database (Denmark)

    Frisvad, Jens Christian; Nielsen, Kristian Fog; Mogensen, Jesper

    Introduction: Fumonisins are toxic seconday metabolites from Fusarium verticillioides and other Fusaria, from Tolypocladium and Aspergillus niger 1,2. Being a generalist Aspergillus niger is the workhorse in a very large number of industrial applications, and is also a common contaminant in foods...... of approximately 6% of the strains. None of the other species in the black Aspergilli produced fumonisins. One strain (NRRL 337), called the “food fungus”, because it is used for single cell protein based on cheap growth substrates, produced both fumonisins and ochratoxin A. Industrial citric acid producers...

  3. Significance and occurrence of fumonisins from Aspergillus niger

    DEFF Research Database (Denmark)

    Mogensen, Jesper Mølgaard

    Fumonisins is a well-studied group of mycotoxins, mainly produced in maize by Fusarium species. However with the recent discovery of a fumonisin production by Aspergillus niger, other food commodities are at risk, since A. niger is a ubiquitous contaminant of many food and feed products. The obje......Fumonisins is a well-studied group of mycotoxins, mainly produced in maize by Fusarium species. However with the recent discovery of a fumonisin production by Aspergillus niger, other food commodities are at risk, since A. niger is a ubiquitous contaminant of many food and feed products....... The objective of this thesis was to determine the significance and occurrence of fumonisins from Aspergillus niger in food, the frequency of fumonisin production in A. niger isolates, as well as studies of the effect of physiological factors affecting fumonisin production. Major findings in this context have...... isolation of 47 black Aspergilli, who all turned out to be A. acidus and did not produce neither ochratoxin or fumonisins, no significant risk from these toxins were associated with this intake. Single kernel analysis of maize showed that fumonisins were present in 15% of the kernels at varying...

  4. A molecular analysis of L-arabinan degradation in Aspergillus niger and Aspergillus nidulans

    NARCIS (Netherlands)

    Flipphi, M.J.A.

    1995-01-01

    This thesis describes a molecular study of the genetics ofL-arabinan degradation in Aspergillus niger and Aspergillus nidulans. These saprophytic hyphal fungi produce an extracellular hydrolytic enzyme system to

  5. Metabolic control analysis of Aspergillus niger L-arabinose catabolism

    NARCIS (Netherlands)

    Groot, de M.J.L.; Prathumpai, W.; Visser, J.; Ruijter, G.J.G.

    2005-01-01

    A mathematical model of the L-arabinose/D-xylose catabolic pathway of Aspergillus niger was constructed based on the kinetic properties of the enzymes. For this purpose L-arabinose reductase, L-arabitol dehydrogenase and D-xylose reductase were purified using dye-affinity chromatography, and their

  6. Biosynthesis of phytases and phosphatases by Aspergillus niger 551

    African Journals Online (AJOL)

    ARyw

    2012-02-16

    Feb 16, 2012 ... Key words: Aspergillus niger, phytase, phosphatase, amylase, protease, starch. ... biological production of phytases is of great significance in research. The researchers focus their attention mainly on finding appropriate strains and on their ... substrate, 200 μl of the enzyme extract and 200 μl 25 mMol of.

  7. Effect Of Laboratory Produced Xylanase From Aspergillus niger On ...

    African Journals Online (AJOL)

    The efficacy of laboratory produced xylanase enzyme from Aspergillus niger on highly fibrous feeds and feedstuffs was investigated. Guinea grass (Panicum maximum) from grass hay and rice husk (Oryza sativa) from crop by-product characterised with high fibre content served as substrates (Feeds and Feedstuffs) for the ...

  8. Comparative study of Aspergillus niger and Penicillium sp. in the ...

    African Journals Online (AJOL)

    The comparative study of Aspergillus niger and Penicillium sp. in the biodegradation of automotive gas oil (AGO) and premium motor spirit (PMS) was carried out to ascertain the effectiveness of using these microorganisms in cleaning and restoring the ecosystem when polluted by petroleum products. These fungi were ...

  9. Cellulase production by wild strains of Aspergillus niger, Penicillium ...

    African Journals Online (AJOL)

    Waste cellulosic materials (corncob, sawdust and sugarcane pulp) and crystalline cellulose induced cellulase production in wild strains of Aspergillus niger, Penicillium chrysogenum and Trichoderma harzianum isolated from a wood-waste dump in Lagos, Nigeria. Cellulose-supplemented media gave the maximum ...

  10. Cellulase and Pectinase Production Potentials of Aspergillus Niger ...

    African Journals Online (AJOL)

    Production of pectinase and cellulase by Aspergillus niger from corn cob was examined. The organism was screened for enzymatic activity using Carboxyl Methyl Cellulose (CMC) and Pectin as substrate. The result revealed a clear zone of inhibition in the agar plates. The organism was subjected to different optimum ...

  11. Genetic relationships among strains of the Aspergillus niger aggregate

    DEFF Research Database (Denmark)

    Ferracin, L.M.; Frisvad, Jens Christian; Taniwaki, M.H.

    2009-01-01

    We analyzed the genetic relationships between 51 fungal isolates previously identified as A. niger aggregate, obtained from dried fruit samples from worldwide origin and 7 A. tubingensis obtained from Brazilian coffee beans samples. Greater fungal diversity was found in black sultanas. Aspergillus...

  12. Production of Aspergillus niger biomass from aqueous extract of ...

    African Journals Online (AJOL)

    single cell protein) production was investigated. Aspergillus niger was grown in unsupplemented and supplemented (glucose or nitrogen sources [(NH4)2SO4, NaNO3, NH4Cl, NH4NO3 and KNO3]) BSG liquor by submerge fermentation on rotary ...

  13. Optimization of chloroxylenol degradation by Aspergillus niger using ...

    African Journals Online (AJOL)

    Chloroxylenol is a very toxic phenolic derivative and it represents potential hazard towards human health and to the environment. Aspergillus niger, local isolate, is an efficient fungus to degrade 99.72% of 2 mg/L of chloroxylenol after 7 days of fermentation. It also has a high capacity to degrade 91.83% of higher ...

  14. Optimization for cellulase production by Aspergillus niger using saw ...

    African Journals Online (AJOL)

    Cellulases are a group of hydrolytic enzymes and are capable of degrading lignocellulosic materials. Cellulases have wide range of applications. This work focuses on factors relevant for improvement of enzymatic hydrolysis of saw dust by using Aspergillus niger. Different cultural conditions were examined to assess their ...

  15. Maturation of conidia on conidiophores of Aspergillus niger

    NARCIS (Netherlands)

    Teertstra, Wieke R.; Tegelaar, Martin; Dijksterhuis, Jan; Golovina, Elena A.; Ohm, Robin A.; Wösten, Han A.B.

    2016-01-01

    Conidia of Aspergillus niger are produced on conidiophores. Here, maturation of conidia on these asexual reproductive structures was studied. Pigmented conidia that had developed on conidiophores for 2, 5, and 8 days were similarly resistant to heat and were metabolically active as shown by CO2

  16. Assessment of Aspergillus niger biofilm growth kinetics in ...

    African Journals Online (AJOL)

    Aspergillus niger ATCC 10864 was grown on bubble minicolumns aerated with pure oxygen either as biofilm or free-mycelium submerged systems. Growth kinetics was followed by estimating biomass from released carbon dioxide using a titration method and mathematical models. Submerged cultures showed increased ...

  17. Comparative study of Aspergillus niger and Penicillium sp. in the ...

    African Journals Online (AJOL)

    STORAGESEVER

    2010-06-14

    Jun 14, 2010 ... The comparative study of Aspergillus niger and Penicillium sp. in the biodegradation of automotive gas oil (AGO) and premium motor spirit (PMS) was carried out to ascertain the effectiveness of using these microorganisms in cleaning and restoring the ecosystem when polluted by petroleum products.

  18. Fumonisin and Ochratoxin Production in Industrial Aspergillus niger Strains

    DEFF Research Database (Denmark)

    Frisvad, Jens Christian; Larsen, Thomas Ostenfeld; Thrane, Ulf

    2011-01-01

    Aspergillus niger is perhaps the most important fungus used in biotechnology, and is also one of the most commonly encountered fungi contaminating foods and feedstuffs, and occurring in soil and indoor environments. Many of its industrial applications have been given GRAS status (generally regard...

  19. Evaluation of xylanases from Aspergillus niger and Trichoderma sp ...

    African Journals Online (AJOL)

    Despite being present in relatively low amounts, pentosans and hemicelluloses play an important role in dough rheology and bread properties. The aim of this work is to understand how the xylanases from Aspergillus niger and Trichoderma sp. influence dough rheology, such as elasticity, extensibility, strength and stability.

  20. Effects Of Solid State Fermentation By Aspergillus niger and ...

    African Journals Online (AJOL)

    fermentation and incubated for O (control), 4,8 and 10 days to evaluate the changes in crude protein (CP), crude fibre (CF), neutral detergent fibre (NDF), acid ... inoculation of Cassava peel meal (CPM) with Aspergillus niger, the crude protein increased from 3.50% to 7.0% and to 5.25% on inoculation with Rhizopus spp.

  1. Metabolic control analysis of Aspergillus niger L-arabinose catabolism

    DEFF Research Database (Denmark)

    de Groot, M.J.L.; Prathumpai, Wai; Visser, J.

    2005-01-01

    A mathematical model of the L-arabinose/D-xylose catabolic pathway of Aspergillus niger was constructed based on the kinetic properties of the enzymes. For this purpose L-arabinose reductase, L-arabitol dehydrogenase and D-xylose reductase were purified using dye-affinity chromatography, and thei...

  2. Mannitol is required for stress tolerance in Aspergillus niger conidiospores

    NARCIS (Netherlands)

    Ruijter, G.J.G.; Bax, M.; Patel, H.; Flitter, S.J.; Vondervoort, van de P.J.I.; Vries, de R.P.; Kuyk, van P.A.; Visser, J.

    2003-01-01

    D-Mannitol is the predominant carbon compound in conidiospores of the filamentous fungus Aspergillus niger and makes up 10 to 15% of the dry weight. A number of physiological functions have been ascribed to mannitol, including serving as a reserve carbon source, as an antioxidant, and to store

  3. Aspergillus niger Secretes Citrate to Increase Iron Bioavailability

    Science.gov (United States)

    Odoni, Dorett I.; van Gaal, Merlijn P.; Schonewille, Tom; Tamayo-Ramos, Juan A.; Martins dos Santos, Vitor A. P.; Suarez-Diez, Maria; Schaap, Peter J.

    2017-01-01

    Aspergillus niger has an innate ability to secrete various organic acids, including citrate. The conditions required for A. niger citrate overproduction are well described, but the physiological reasons underlying extracellular citrate accumulation are not yet fully understood. One of the less understood culture conditions is the requirement of growth-limiting iron concentrations. While this has been attributed to iron-dependent citrate metabolizing enzymes, this straightforward relationship does not always hold true. Here, we show that an increase in citrate secretion under iron limited conditions is a physiological response consistent with a role of citrate as A. niger iron siderophore. We found that A. niger citrate secretion increases with decreasing amounts of iron added to the culture medium and, in contrast to previous findings, this response is independent of the nitrogen source. Differential transcriptomics analyses of the two A. niger mutants NW305 (gluconate non-producer) and NW186 (gluconate and oxalate non-producer) revealed up-regulation of the citrate biosynthesis gene citA under iron limited conditions compared to iron replete conditions. In addition, we show that A. niger can utilize Fe(III) citrate as iron source. Finally, we discuss our findings in the general context of the pH-dependency of A. niger organic acid production, offering an explanation, besides competition, for why A. niger organic acid production is a sequential process influenced by the external pH of the culture medium. PMID:28824560

  4. Contribution of arginase to manganese metabolism of Aspergillus niger.

    Science.gov (United States)

    Keni, Sarita; Punekar, Narayan S

    2016-02-01

    Aspects of manganese metabolism during normal and acidogenic growth of Aspergillus niger were explored. Arginase from this fungus was a Mn[II]-enzyme. The contribution of the arginase protein towards A. niger manganese metabolism was investigated using arginase knockout (D-42) and arginase over-expressing (ΔXCA-29) strains of A. niger NCIM 565. The Mn[II] contents of various mycelial fractions were found in the order: D-42 strain niger mycelia harvested from acidogenic growth media contain substantially less Mn[II] as compared to those from normal growth media. Nevertheless, acidogenic mycelia harbor considerable Mn[II] levels and a functional arginase. Altered levels of mycelial arginase protein did not significantly influence citric acid production. The relevance of arginase to cellular Mn[II] pool and homeostasis was evaluated and the results suggest that arginase regulation could occur via manganese availability.

  5. Aspergillus tubingensis and Aspergillus niger as the dominant black Aspergillus, use of simple PCR-RFLP for preliminary differentiation.

    Science.gov (United States)

    Mirhendi, H; Zarei, F; Motamedi, M; Nouripour-Sisakht, S

    2016-03-01

    This work aimed to identify the species distribution of common clinical and environmental isolates of black Aspergilli based on simple restriction fragment length polymorphism (RFLP) analysis of the β-tubulin gene. A total of 149 clinical and environmental strains of black Aspergilli were collected and subjected to preliminary morphological examination. Total genomic DNAs were extracted, and PCR was performed to amplify part of the β-tubulin gene. At first, 52 randomly selected samples were species-delineated by sequence analysis. In order to distinguish the most common species, PCR amplicons of 117 black Aspergillus strains were identified by simple PCR-RFLP analysis using the enzyme TasI. Among 52 sequenced isolates, 28 were Aspergillus tubingensis, 21 Aspergillus niger, and the three remaining isolates included Aspergillus uvarum, Aspergillus awamori, and Aspergillus acidus. All 100 environmental and 17 BAL samples subjected to TasI-RFLP analysis of the β-tubulin gene, fell into two groups, consisting of about 59% (n=69) A. tubingensis and 41% (n=48) A. niger. Therefore, the method successfully and rapidly distinguished A. tubingensis and A. niger as the most common species among the clinical and environmental isolates. Although tardy, the Ehrlich test was also able to differentiate A. tubingensis and A. niger according to the yellow color reaction specific to A. niger. A. tubingensis and A. niger are the most common black Aspergillus in both clinical and environmental isolates in Iran. PCR-RFLP using TasI digestion of β-tubulin DNA enables rapid screening for these common species. Copyright © 2016 Elsevier Masson SAS. All rights reserved.

  6. Cadmium biosorption by Aspergillus niger; Biossorcao de cadmio pelo Aspergillus niger

    Energy Technology Data Exchange (ETDEWEB)

    Silva, E.P.; Barros Junior, L.M.; Duarte, M.M.L.; Macedo, G.R. [Universidade Federal do Rio Grande do Norte, Natal, RN (Brazil)]. E-mail: edmilson@eq.ufrn.br

    2003-07-01

    Biosorption is a property of certain types of inactive, dead, microbial biomass to bind and concentrate heavy metals from even very dilute aqueous solutions. Biomass exhibits this property, acting just as a chemical substance, as an ion exchanger of biological origin. It is particularly the cell wall structure of certain algae, fungi and bacteria which was found responsible for this phenomenon. Some of the biomass types come as a waste by-product of large-scale industrial fermentations (the mold Rhizopus or the bacterium Bacillus subtilis). Other metal-binding biomass types, certain abundant seaweeds (particularly brown algae e.g. Sargassum, Ecklonia), can be readily collected from the oceans. These biomass types, serving as a basis for metal biosorption processes, can accumulate in excess of 25% of their dry weight in deposited heavy metals: Pb, Cd, U, Cu, Zn, even Cr and others. Sorption experiments using the Aspergillus niger fungus for cadmium removal were carried out to study the factors influencing and optimizing the biosorption of this metal. The effects of pH, time, biomass concentration, and initial concentration of the heavy metal on the rate of metallic biosorption were examined. (author)

  7. Intra and extracellular nuclease production by Aspergillus niger and Aspergillus nidulans

    Directory of Open Access Journals (Sweden)

    Ferreira Adlane V. B.

    1998-01-01

    Full Text Available Intra and extracellular nuclease production by strains of Aspergillus niger and Aspergillus nidulans was estimated using a modified DNAse test agar and cell-free extract assays. Differences in the production of nucleases by A. niger and A. nidulans were observed. These observations suggest that the DNAse test agar can be helpful for a quick screening for some types of nucleases in filamentous fungi. The assays using cell-free extracts can also be useful for initial characterization of other types of nucleases.

  8. Production of extremophilic bacterial cellulase enzymes in aspergillus niger.

    Energy Technology Data Exchange (ETDEWEB)

    Gladden, John Michael

    2013-09-01

    Enzymes can be used to catalyze a myriad of chemical reactions and are a cornerstone in the biotechnology industry. Enzymes have a wide range of uses, ranging from medicine with the production of pharmaceuticals to energy were they are applied to biofuel production. However, it is difficult to produce large quantities of enzymes, especially if they are non-native to the production host. Fortunately, filamentous fungi, such as Aspergillus niger, are broadly used in industry and show great potential for use a heterologous enzyme production hosts. Here, we present work outlining an effort to engineer A. niger to produce thermophilic bacterial cellulases relevant to lignocellulosic biofuel production.

  9. Recombinant bacterial hemoglobin alters metabolism of Aspergillus niger

    DEFF Research Database (Denmark)

    Hofmann, Gerald; Diano, Audrey; Nielsen, Jens

    2009-01-01

    The filamentous fungus Aspergillus niger is used extensively for the production of enzymes and organic acids. A major problem in industrial fermentations with this fungus is to ensure sufficient supply of oxygen required for respiratory metabolism of the fungus. In case of oxygen limitation...... behind the strong gpdA promoter from Aspergillus nidulans. Analysis of secreted metabolites, oxygen uptake, CO2 evolution and biomass formation points towards a relief of stress in the mutant expressing VHB when it is exposed to oxygen limitation. Our findings therefore point to an interesting strategy...

  10. The potential of Aspergillus fumigatus and Aspergillus niger in ...

    African Journals Online (AJOL)

    SARAH

    2015-10-31

    :1, 1:3 and 1:5 respectively for. 3 weeks. ... ability of A. fumigatus and A. niger to bioaccumulate less toxic metals such as Iron, Lead and Zinc at high concentrations in the ... ability to remediate industrial wastewater containing.

  11. Hypersensitivity testing for Aspergillus fumigatus IgE is significantly more sensitive than testing for Aspergillus niger IgE.

    Science.gov (United States)

    Selvaggi, Thomas A; Walco, Jeremy P; Parikh, Sujal; Walco, Gary A

    2012-02-01

    We sought to determine if sufficient redundancy exists between specific IgE testing for Aspergillus fumigatus and Aspergillus niger to eliminate one of the assays in determining Aspergillus hypersensitivity. We reviewed regional laboratory results comparing A fumigatus-specific IgE with A niger-specific IgE using the Pharmacia UniCAP system (Pharmacia, Kalamazoo, MI). By using the Fisher exact test as an index of concordance among paired results, we showed a significant difference between 109 paired samples for the presence of specific IgE to A fumigatus and A niger (P niger; no specimen was positive for A niger and negative for A fumigatus. We conclude that A fumigatus-specific IgE is sufficient to detect Aspergillus hypersensitivity. The assay for A niger-specific IgE is redundant, less sensitive, and unnecessary if the assay for specific IgE for A fumigatus is performed.

  12. Biosynthesis of phytases and phosphatases by Aspergillus niger 551

    African Journals Online (AJOL)

    The purpose of this study was to determine the ability of the Aspergillus niger 551 strain to produce phytases and phosphatases in 24 days shake cultures at a pH adjusted to 5.5 and a changing (uncontrolled) pH, in a medium containing 0.5% of glucose and 4% of starch. The fermentation process of the culture performed ...

  13. Optimization of citric acid production by Aspergillus niger

    OpenAIRE

    H.R Samadlouie; Sh Gharanjik

    2015-01-01

    Among the various fungal strains screened for citric acid production, Aspergillus niger is known to produce considerable amounts of citric acid and other organic acids when cultivated in submerge fermentation. In this study, optimization of the medium components was carried out using "one-factor-at-a-time" and response surface methods (RSM). One-factor-at-a-time indicated that the amount of citric acid production was increased along with the increasing of agitation speed (from 150 to 200 rpm)...

  14. Growth and hydrolase profiles can be used as characteristics to distinguish Aspergillus niger and other black aspergilli

    Science.gov (United States)

    Meijer, M.; Houbraken, J.A.M.P.; Dalhuijsen, S.; Samson, R.A.; de Vries, R.P.

    2011-01-01

    Wild type Aspergillus niger isolates from different biotopes from all over the world were compared to each other and to the type strains of other black Aspergillus species with respect to growth and extracellular enzyme profiles. The origin of the A. niger isolate did not result in differences in growth profile with respect to monomeric or polymeric carbon sources. Differences were observed in the growth rate of the A. niger isolates, but these were observed on all carbon sources and not specific for a particular carbon source. In contrast, carbon source specific differences were observed between the different species. Aspergillus brasiliensis is the only species able to grow on D-galactose, and A. aculeatus had significantly better growth on Locus Bean gum than the other species. Only small differences were found in the extracellular enzyme profile of the A. niger isolates during growth on wheat bran, while large differences were observed in the profiles of the different black aspergilli. In addition, differences were observed in temperature profiles between the black Aspergillus species, but not between the A. niger isolates, demonstrating no isolate-specific adaptations to the environment. These data indicate that the local environment does not result in stable adaptations of A. niger with respect to growth profile or enzyme production, but that the potential is maintained irrespective of the environmental parameters. It also demonstrates that growth, extracellular protein and temperature profiles can be used for species identification within the group of black aspergilli. PMID:21892240

  15. Production and Purification of Peroxidase from Aspergillus niger.

    Directory of Open Access Journals (Sweden)

    Mohammed A. Jebor

    2017-02-01

    Full Text Available This study was conducted in the laboratories of Biology Department, College of Science, which deals with isolation and purification of peroxidase and optimization of process parameters to achieve maximum yield of peroxidase by Aspergillus niger. Solid-state fermentation of Aspergillus niger was carried out for enhanced production of peroxidase using hydrogen peroxide as the substrate of enzyme maximum activity of the enzyme was achieved under optimum growth conditions. The optimum conditions were the isolated of Aspergillus niger from soil and growth in synthetic medium, it gave high titer of peroxidase activity, the fructose as carbon source, peptone as nitrogen source, after 12 days of incubation, incubation temperature 25 °C and pH = 6.5. Peroxidase purified in four purification steps; precipitation with 70% saturation of ammonium sulfate, step of dialysis, the third by ion exchange chromatography using DEAE-Cellulose and fourth by gel filtration throughout Sephadex G-100. The specific activity of the purified enzyme was 150U/mg with 7.75 folds. The peroxidase was shown to have molecular weight of 40kDa in SDS-PAGA and about 40kDa in gel filtration.The optimum pH and temperature for peroxidase activity 7 and 35 C0 respectively.

  16. Physicochemical Properties Analysis and Secretome of Aspergillus niger in Fermented Rapeseed Meal

    National Research Council Canada - National Science Library

    Shi, Changyou; He, Jun; Yu, Jie; Yu, Bing; Mao, Xiangbing; Zheng, Ping; Huang, Zhiqing; Chen, Daiwen

    2016-01-01

    .... To further understand possible mechanism of solid state fermentation, the composition of extracellular enzymes secreted by Aspergillus niger during fermentation was analysed using two-dimentional...

  17. Novel Route for Agmatine Catabolism in Aspergillus niger Involves 4-Guanidinobutyrase.

    Science.gov (United States)

    Kumar, Sunil; Saragadam, Tejaswani; Punekar, Narayan S

    2015-08-15

    Agmatine, a significant polyamine in bacteria and plants, mostly arises from the decarboxylation of arginine. The functional importance of agmatine in fungi is poorly understood. The metabolism of agmatine and related guanidinium group-containing compounds in Aspergillus niger was explored through growth, metabolite, and enzyme studies. The fungus was able to metabolize and grow on l-arginine, agmatine, or 4-guanidinobutyrate as the sole nitrogen source. Whereas arginase defined the only route for arginine catabolism, biochemical and bioinformatics approaches suggested the absence of arginine decarboxylase in A. niger. Efficient utilization by the parent strain and also by its arginase knockout implied an arginase-independent catabolic route for agmatine. Urea and 4-guanidinobutyrate were detected in the spent medium during growth on agmatine. The agmatine-grown A. niger mycelia contained significant levels of amine oxidase, 4-guanidinobutyraldehyde dehydrogenase, 4-guanidinobutyrase (GBase), and succinic semialdehyde dehydrogenase, but no agmatinase activity was detected. Taken together, the results support a novel route for agmatine utilization in A. niger. The catabolism of agmatine by way of 4-guanidinobutyrate to 4-aminobutyrate into the Krebs cycle is the first report of such a pathway in any organism. A. niger GBase peptide fragments were identified by tandem mass spectrometry analysis. The corresponding open reading frame from the A. niger NCIM 565 genome was located and cloned. Subsequent expression of GBase in both Escherichia coli and A. niger along with its disruption in A. niger functionally defined the GBase locus (gbu) in the A. niger genome. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

  18. Expression of the Aspergillus terreus itaconic acid biosynthesis cluster in Aspergillus niger.

    Science.gov (United States)

    van der Straat, Laura; Vernooij, Marloes; Lammers, Marieke; van den Berg, Willy; Schonewille, Tom; Cordewener, Jan; van der Meer, Ingrid; Koops, Andries; de Graaff, Leo H

    2014-01-17

    Aspergillus terreus is a natural producer of itaconic acid and is currently used to produce itaconic acid on an industrial scale. The metabolic process for itaconic acid biosynthesis is very similar to the production of citric acid in Aspergillus niger. However, a key enzyme in A. niger, cis-aconitate decarboxylase, is missing. The introduction of the A. terreus cadA gene in A. niger exploits the high level of citric acid production (over 200 g per liter) and theoretically can lead to production levels of over 135 g per liter of itaconic acid in A. niger. Given the potential for higher production levels in A. niger, production of itaconic acid in this host was investigated. Expression of Aspergillus terreus cis-aconitate decarboxylase in Aspergillus niger resulted in the production of a low concentration (0.05 g/L) of itaconic acid. Overexpression of codon-optimized genes for cis-aconitate decarboxylase, a mitochondrial transporter and a plasma membrane transporter in an oxaloacetate hydrolase and glucose oxidase deficient A. niger strain led to highly increased yields and itaconic acid production titers. At these higher production titers, the effect of the mitochondrial and plasma membrane transporters was much more pronounced, with levels being 5-8 times higher than previously described. Itaconic acid can be produced in A. niger by the introduction of the A. terreus cis-aconitate decarboxylase encoding cadA gene. This results in a low itaconic acid production level, which can be increased by codon-optimization of the cadA gene for A. niger. A second crucial requirement for efficient production of itaconic acid is the expression of the A. terreus mttA gene, encoding a putative mitochondrial transporter. Expression of this transporter results in a twenty-fold increase in the secretion of itaconic acid. Expression of the A. terreus itaconic acid cluster consisting of the cadA gene, the mttA gene and the mfsA gene results in A. niger strains that produce over

  19. Biosorption of Pentachlorophenol from Aqueous Solutions by Aspergillus Niger Biomass

    Directory of Open Access Journals (Sweden)

    Reza Shokoohi

    2016-06-01

    Full Text Available Background: This study aimed to investigate the biosorption of pentachlorophenol on Aspergillus niger biomass as a method for removal of pentachlorophenol from aqueous solutions. Methods: Modified A. niger biomass with NaOH was used to absorb the pentachlorophenol. The impacts of various experimental parameters like primary pentachlorophenol concentration, pH of the solution, contact time, and biomass dosage on the biosorption of pentachlorophenol were investigated. Results: The correlation of contact time, pH and initial concentration with the biosorption of pentachlorophenol by A. niger biomass was statistically significant (P<0.001. Pentachlorophenol removal increased with decreasing pH of the solution and the maximum efficiency was obtained at pH=3. The equilibrium adsorption capacity was increased from 4.23 to 11.65 mg/g by increasing initial pentachlorophenol concentration from 10 to 40 mg/L, while pentachlorophenol removal efficiency decreased from 87 to 55%. Both Langmuir and Freundlich isotherms efficiently described adsorption equilibrium of pentachlorophenol on A. niger biomass. Correlation coefficients for the second order kinetic model were almost equal to one. Conclusion: A. niger biomass can be used to reduce the toxicity of aqueous solutions containing pentachlorophenol in acidic pH conditions.

  20. Transcriptome analysis of Aspergillus niger grown on sugarcane bagasse

    Science.gov (United States)

    2011-01-01

    Background Considering that the costs of cellulases and hemicellulases contribute substantially to the price of bioethanol, new studies aimed at understanding and improving cellulase efficiency and productivity are of paramount importance. Aspergillus niger has been shown to produce a wide spectrum of polysaccharide hydrolytic enzymes. To understand how to improve enzymatic cocktails that can hydrolyze pretreated sugarcane bagasse, we used a genomics approach to investigate which genes and pathways are transcriptionally modulated during growth of A. niger on steam-exploded sugarcane bagasse (SEB). Results Herein we report the main cellulase- and hemicellulase-encoding genes with increased expression during growth on SEB. We also sought to determine whether the mRNA accumulation of several SEB-induced genes encoding putative transporters is induced by xylose and dependent on glucose. We identified 18 (58% of A. niger predicted cellulases) and 21 (58% of A. niger predicted hemicellulases) cellulase- and hemicellulase-encoding genes, respectively, that were highly expressed during growth on SEB. Conclusions Degradation of sugarcane bagasse requires production of many different enzymes which are regulated by the type and complexity of the available substrate. Our presently reported work opens new possibilities for understanding sugarcane biomass saccharification by A. niger hydrolases and for the construction of more efficient enzymatic cocktails for second-generation bioethanol. PMID:22008461

  1. Transcriptome analysis of Aspergillus niger grown on sugarcane bagasse

    Directory of Open Access Journals (Sweden)

    Goldman Gustavo H

    2011-10-01

    Full Text Available Abstract Background Considering that the costs of cellulases and hemicellulases contribute substantially to the price of bioethanol, new studies aimed at understanding and improving cellulase efficiency and productivity are of paramount importance. Aspergillus niger has been shown to produce a wide spectrum of polysaccharide hydrolytic enzymes. To understand how to improve enzymatic cocktails that can hydrolyze pretreated sugarcane bagasse, we used a genomics approach to investigate which genes and pathways are transcriptionally modulated during growth of A. niger on steam-exploded sugarcane bagasse (SEB. Results Herein we report the main cellulase- and hemicellulase-encoding genes with increased expression during growth on SEB. We also sought to determine whether the mRNA accumulation of several SEB-induced genes encoding putative transporters is induced by xylose and dependent on glucose. We identified 18 (58% of A. niger predicted cellulases and 21 (58% of A. niger predicted hemicellulases cellulase- and hemicellulase-encoding genes, respectively, that were highly expressed during growth on SEB. Conclusions Degradation of sugarcane bagasse requires production of many different enzymes which are regulated by the type and complexity of the available substrate. Our presently reported work opens new possibilities for understanding sugarcane biomass saccharification by A. niger hydrolases and for the construction of more efficient enzymatic cocktails for second-generation bioethanol.

  2. Fumonisin and Ochratoxin Production in Industrial Aspergillus niger Strains

    Science.gov (United States)

    Frisvad, Jens C.; Larsen, Thomas O.; Thrane, Ulf; Meijer, Martin; Varga, Janos; Samson, Robert A.; Nielsen, Kristian F.

    2011-01-01

    Aspergillus niger is perhaps the most important fungus used in biotechnology, and is also one of the most commonly encountered fungi contaminating foods and feedstuffs, and occurring in soil and indoor environments. Many of its industrial applications have been given GRAS status (generally regarded as safe). However, A. niger has the potential to produce two groups of potentially carcinogenic mycotoxins: fumonisins and ochratoxins. In this study all available industrial and many non-industrial strains of A. niger (180 strains) as well as 228 strains from 17 related black Aspergillus species were examined for mycotoxin production. None of the related 17 species of black Aspergilli produced fumonisins. Fumonisins (B2, B4, and B6) were detected in 81% of A. niger, and ochratoxin A in 17%, while 10% of the strains produced both mycotoxins. Among the industrial strains the same ratios were 83%, 33% and 26% respectively. Some of the most frequently used strains in industry NRRL 337, 3112 and 3122 produced both toxins and several strains used for citric acid production were among the best producers of fumonisins in pure agar culture. Most strains used for other biotechnological processes also produced fumonisins. Strains optimized through random mutagenesis usually maintained their mycotoxin production capability. Toxigenic strains were also able to produce the toxins on media suggested for citric acid production with most of the toxins found in the biomass, thereby questioning the use of the remaining biomass as animal feed. In conclusion it is recommended to use strains of A. niger with inactive or inactivated gene clusters for fumonisins and ochratoxins, or to choose isolates for biotechnological uses in related non-toxigenic species such as A. tubingensis, A. brasiliensis, A vadensis or A. acidus, which neither produce fumonisins nor ochratoxins. PMID:21853139

  3. Fumonisin and ochratoxin production in industrial Aspergillus niger strains.

    Directory of Open Access Journals (Sweden)

    Jens C Frisvad

    Full Text Available Aspergillus niger is perhaps the most important fungus used in biotechnology, and is also one of the most commonly encountered fungi contaminating foods and feedstuffs, and occurring in soil and indoor environments. Many of its industrial applications have been given GRAS status (generally regarded as safe. However, A. niger has the potential to produce two groups of potentially carcinogenic mycotoxins: fumonisins and ochratoxins. In this study all available industrial and many non-industrial strains of A. niger (180 strains as well as 228 strains from 17 related black Aspergillus species were examined for mycotoxin production. None of the related 17 species of black Aspergilli produced fumonisins. Fumonisins (B(2, B(4, and B(6 were detected in 81% of A. niger, and ochratoxin A in 17%, while 10% of the strains produced both mycotoxins. Among the industrial strains the same ratios were 83%, 33% and 26% respectively. Some of the most frequently used strains in industry NRRL 337, 3112 and 3122 produced both toxins and several strains used for citric acid production were among the best producers of fumonisins in pure agar culture. Most strains used for other biotechnological processes also produced fumonisins. Strains optimized through random mutagenesis usually maintained their mycotoxin production capability. Toxigenic strains were also able to produce the toxins on media suggested for citric acid production with most of the toxins found in the biomass, thereby questioning the use of the remaining biomass as animal feed. In conclusion it is recommended to use strains of A. niger with inactive or inactivated gene clusters for fumonisins and ochratoxins, or to choose isolates for biotechnological uses in related non-toxigenic species such as A. tubingensis, A. brasiliensis, A vadensis or A. acidus, which neither produce fumonisins nor ochratoxins.

  4. D-Galactose uptake is nonfunctional in the conidiospores of Aspergillus niger

    NARCIS (Netherlands)

    Fekete, E.; de Vries, R.P.; Seiboth, B.; vanKuyk, P.A.; Sandor, E.; Metz, B.; Kubicek, C.P.; Karaffa, L.

    2012-01-01

    The majority of black Aspergilli (Aspergillus section Nigri), including Aspergillus niger, as well as many other Ascomycetes fail to germinate on d-galactose as a sole carbon source. Here, we provide evidence that the ability of A. niger to transport d-galactose is growth stage dependent, being

  5. Reduced by-product formation and modified oxygen availability improve itaconic acid production in Aspergillus niger.

    NARCIS (Netherlands)

    Li, A.; Pfelzer, N.; Zuijderwijk, R.; Brickwedde, A.; Zeijl, C. van; Punt, P.

    2013-01-01

    Aspergillus niger has an extraordinary potential to produce organic acids as proven by its application in industrial citric acid production. Previously, it was shown that expression of the cis-aconitate decarboxylase gene (cadA) from Aspergillus terreus converted A. niger into an itaconic acid

  6. Engineering of Aspergillus niger for the production of secondary metabolites.

    Science.gov (United States)

    Richter, Lennart; Wanka, Franziska; Boecker, Simon; Storm, Dirk; Kurt, Tutku; Vural, Özlem; Süßmuth, Roderich; Meyer, Vera

    2014-01-01

    Filamentous fungi can each produce dozens of secondary metabolites which are attractive as therapeutics, drugs, antimicrobials, flavour compounds and other high-value chemicals. Furthermore, they can be used as an expression system for eukaryotic proteins. Application of most fungal secondary metabolites is, however, so far hampered by the lack of suitable fermentation protocols for the producing strain and/or by low product titers. To overcome these limitations, we report here the engineering of the industrial fungus Aspergillus niger to produce high titers (up to 4,500 mg • l-1) of secondary metabolites belonging to the class of nonribosomal peptides. For a proof-of-concept study, we heterologously expressed the 351 kDa nonribosomal peptide synthetase ESYN from Fusarium oxysporum in A. niger. ESYN catalyzes the formation of cyclic depsipeptides of the enniatin family, which exhibit antimicrobial, antiviral and anticancer activities. The encoding gene esyn1 was put under control of a tunable bacterial-fungal hybrid promoter (Tet-on) which was switched on during early-exponential growth phase of A. niger cultures. The enniatins were isolated and purified by means of reverse phase chromatography and their identity and purity proven by tandem MS, NMR spectroscopy and X-ray crystallography. The initial yields of 1 mg • l-1 of enniatin were increased about 950 fold by optimizing feeding conditions and the morphology of A. niger in liquid shake flask cultures. Further yield optimization (about 4.5 fold) was accomplished by cultivating A. niger in 5 l fed batch fermentations. Finally, an autonomous A. niger expression host was established, which was independent from feeding with the enniatin precursor d-2-hydroxyvaleric acid d-Hiv. This was achieved by constitutively expressing a fungal d-Hiv dehydrogenase in the esyn1-expressing A. niger strain, which used the intracellular α-ketovaleric acid pool to generate d-Hiv. This is the first report demonstrating that A

  7. Optimization of cellulase production by Aspergillus niger NCIM 1207.

    Science.gov (United States)

    Gokhale, D V; Patil, S G; Bastawde, K B

    1991-07-01

    Aspergillus niger NCIM 1207 produces high levels of extracellular beta-glucosidase and xylanase activities in submerged fermentation. Among the nitrogen sources, ammonium sulfate, ammonium dihydrogen orthophosphate, and corn-steep liquor were the best for the production of cellulolytic enzymes by A. niger. The optimum pH and temperature for cellulase production were 3.0-5.5 and 28 degrees C, respectively. The cellulase complex of this strain was found to undergo catabolite repression in the presence of high concentrations of glucose. Glycerol at all concentrations caused catabolite repression of cellulase production. The addition of glucose (up to 1% concentration) enhanced the production of cellulolytic enzymes, but a higher concentration of glucose effected the pronounced repression of enzymes. Generally the growth on glucose- or glycerol-containing medium was accompanied by a sudden drop in the pH of the fermentation medium to 2.0.

  8. Production of Citric Acid by Aspergillus niger Using Pineapple Waste

    Directory of Open Access Journals (Sweden)

    Kareem, S. O.

    2010-01-01

    Full Text Available A solid state fermentation was developed for citric acid production from pineapple waste by Aspergillus niger KS-7. The medium was supplemented with different concentration of glucose, sucrose, ammonium nitrate and ammonium phosphate. It was found that pineapple waste with 15% (w/v sucrose and ammonium nitrate (0.25% w/v gave the optimum citric acid secretion (60.61 g/kg in the presence of methanol (2% v/v when fermented for 5 days at 30 °C with the initial moisture content of 65%. The yield was more than 90% based on the amount of fermentable sugar consumed. These results present the use of pineapple peel as a cheap medium for the production of commercially valuable organic acid by A. niger.

  9. The Effectiveness of Antifungal Controlling Aspergillus Niger Growth on Plasterboard

    Directory of Open Access Journals (Sweden)

    Parjo Umi Kalthsom

    2017-01-01

    Full Text Available Good indoor environmental quality is desired for a healthy indoor environment. The microbial growth under indoor environments contribute to the poor indoor environmental quality that can cause various of health problems. In this study, the applications of three types of antifungals to prevent microbial migration, subsequent growth and bio-deterioration of the substrates. The aim of this research was to evaluate the coating-bio resistance in remediation of indoor fungal using three types of antifungals with different types of wall finishing materials. The treatment was exposed to optimum temperature and relative humidity at 30°C and 90% respectively. The potassium sorbate, zinc salicylate and calcium benzoate are tested against Aspergillus niger which is collected from indoor rooms. This study has revealed the growth of A. niger are more affected by the potassium sorbate on thick wallpaper, which is the percentage growth are 47%.

  10. FluG affects secretion in colonies of Aspergillus niger.

    Science.gov (United States)

    Wang, Fengfeng; Krijgsheld, Pauline; Hulsman, Marc; de Bekker, Charissa; Müller, Wally H; Reinders, Marcel; de Vries, Ronald P; Wösten, Han A B

    2015-01-01

    Colonies of Aspergillus niger are characterized by zonal heterogeneity in growth, sporulation, gene expression and secretion. For instance, the glucoamylase gene glaA is more highly expressed at the periphery of colonies when compared to the center. As a consequence, its encoded protein GlaA is mainly secreted at the outer part of the colony. Here, multiple copies of amyR were introduced in A. niger. Most transformants over-expressing this regulatory gene of amylolytic genes still displayed heterogeneous glaA expression and GlaA secretion. However, heterogeneity was abolished in transformant UU-A001.13 by expressing glaA and secreting GlaA throughout the mycelium. Sequencing the genome of UU-A001.13 revealed that transformation had been accompanied by deletion of part of the fluG gene and disrupting its 3' end by integration of a transformation vector. Inactivation of fluG in the wild-type background of A. niger also resulted in breakdown of starch under the whole colony. Asexual development of the ∆fluG strain was not affected, unlike what was previously shown in Aspergillus nidulans. Genes encoding proteins with a signal sequence for secretion, including part of the amylolytic genes, were more often downregulated in the central zone of maltose-grown ∆fluG colonies and upregulated in the intermediate part and periphery when compared to the wild-type. Together, these data indicate that FluG of A. niger is a repressor of secretion.

  11. Adsorption of indigo carmine on dead biomass of aspergillus niger

    OpenAIRE

    Chaves, Karina Oliveira; Monteiro, Carlo Rannyêr Lopes; Muniz, Celli Rodrigues; Gomes, Raimundo Bemvindo; Buarque, Hugo Leonardo de Brito

    2008-01-01

    As indústrias têxteis geram efluentes ricos em corantes durante toda a sua cadeia produtiva. Neste estudo, ensaios cinéticos foram realizados utilizando biomassas mortas de Aspergillus niger, diferentemente tratadas, como adsorvente para a remoção de índigo carmim a partir de soluções aquosas. Ensaio cinético com carvão ativado comercial também foi realizado para comparação. Os resultados obtidos indicam que a biomassa fúngica sem tratamento algum foi capaz de remover 95% do índigo carmim da ...

  12. Antibiotic Extraction as a Recent Biocontrol Method for Aspergillus Niger andAspergillus Flavus Fungi in Ancient Egyptian mural paintings

    Science.gov (United States)

    Hemdan, R. Elmitwalli; Fatma, Helmi M.; Rizk, Mohammed A.; Hagrassy, Abeer F.

    Biodeterioration of mural paintings by Aspergillus niger and Aspergillus flavus Fungi has been proved in different mural paintings in Egypt nowadays. Several researches have studied the effect of fungi on mural paintings, the mechanism of interaction and methods of control. But none of these researches gives us the solution without causing a side effect. In this paper, for the first time, a recent treatment by antibiotic "6 penthyl α pyrone phenol" was applied as a successful technique for elimination of Aspergillus niger and Aspergillus flavus. On the other hand, it is favorable for cleaning Surfaces of Murals executed by tembera technique from the fungi metabolism which caused a black pigments on surfaces.

  13. Post-genomic insights into the plant polysaccharide degradation potential of Aspergillus nidulans and comparison to Aspergillus niger and Aspergillus oryzae

    NARCIS (Netherlands)

    Coutinho, Pedro M; Andersen, Mikael R; Kolenova, Katarina; vanKuyk, Patricia A; Benoit, Isabelle; Gruben, Birgit S; Trejo-Aguilar, Blanca; Visser, Hans; van Solingen, Piet; Pakula, Tiina; Seiboth, Bernard; Battaglia, Evy; Aguilar-Osorio, Guillermo; de Jong, Jan F; Ohm, Robin A; Aguilar, Mariana; Henrissat, Bernard; Nielsen, Jens; Stålbrand, Henrik; de Vries, Ronald P

    The plant polysaccharide degradative potential of Aspergillus nidulans was analysed in detail and compared to that of Aspergillus niger and Aspergillus oryzae using a combination of bioinformatics, physiology and transcriptomics. Manual verification indicated that 28.4% of the A. nidulans ORFs

  14. Molecular Dynamics Approach in Designing Thermostable Aspergillus niger Xylanase

    Science.gov (United States)

    Malau, N. D.; Sianturi, M.

    2017-03-01

    Molecular dynamics methods we have applied as a tool in designing thermostable Aspergillus niger Xylanase, by examining Root Mean Square Deviation (RMSD) and The Stability of the Secondary Structure of enzymes structure at its optimum temperature and compare with its high temperature behavior. As RMSD represents structural fluctuation at a particular temperature, a better understanding of this factor will suggest approaches to bioengineer these enzymes to enhance their thermostability. In this work molecular dynamic simulations of Aspergillus niger xylanase (ANX) have been carried at 400K (optimum catalytic temperature) for 2.5 ns and 500K (ANX reported inactive temperature) for 2.5 ns. Analysis have shown that the Root Mean Square Deviation (RMSD) significant increase at higher temperatures compared at optimum temperature and some of the secondary structures of ANX that have been damaged at high temperature. Structural analysis revealed that the fluctuations of the α-helix and β-sheet regions are larger at higher temperatures compared to the fluctuations at optimum temperature.

  15. An antifungal role of hydrogen sulfide on the postharvest pathogens Aspergillus niger and Penicillium italicum.

    Directory of Open Access Journals (Sweden)

    Liu-Hui Fu

    Full Text Available In this research, the antifungal role of hydrogen sulfide (H2S on the postharvest pathogens Aspergillus niger and Penicillium italicum growing on fruits and under culture conditions on defined media was investigated. Our results show that H2S, released by sodium hydrosulfide (NaHS effectively reduced the postharvest decay of fruits induced by A. niger and P. italicum. Furthermore, H2S inhibited spore germination, germ tube elongation, mycelial growth, and produced abnormal mycelial contractions when the fungi were grown on defined media in Petri plates. Further studies showed that H2S could cause an increase in intracellular reactive oxygen species (ROS in A. niger. In accordance with this observation we show that enzyme activities and the expression of superoxide dismutase (SOD and catalase (CAT genes in A. niger treated with H2S were lower than those in control. Moreover, H2S also significantly inhibited the growth of Saccharomyces cerevisiae, Rhizopus oryzae, the human pathogen Candida albicans, and several food-borne bacteria. We also found that short time exposure of H2S showed a microbicidal role rather than just inhibiting the growth of microbes. Taken together, this study suggests the potential value of H2S in reducing postharvest loss and food spoilage caused by microbe propagation.

  16. Improving cellulase production by Aspergillus niger using adaptive evolution.

    Science.gov (United States)

    Patyshakuliyeva, Aleksandrina; Arentshorst, Mark; Allijn, Iris E; Ram, Arthur F J; de Vries, Ronald P; Gelber, Isabelle Benoit

    2016-06-01

    To evaluate the potential of adaptive evolution as a tool in generating strains with an improved production of plant biomass degrading enzymes. An Aspergillus niger cellulase mutant was obtained by adaptive evolution. Physiological properties of this mutant revealed a five times higher cellulose production than the parental strain. Transcriptomic analysis revealed that the expression of noxR, encoding the regulatory subunit of the NADPH oxidase complex, was reduced in the mutant compared to the parental strain. Subsequent analysis of a noxR knockout strain showed the same phenotypic effect as observed for the evolution mutant, confirming the role of NoxR in cellulose degradation. Adaptive evolution is an efficient approach to modify a strain and activate genes involved in polysaccharide degradation.

  17. In-silico analysis of Aspergillus niger beta-glucosidases

    Science.gov (United States)

    Yeo S., L.; Shazilah, K.; Suhaila, S.; Abu Bakar F., D.; Murad A. M., A.

    2014-09-01

    Genomic data mining was carried out and revealed a total of seventeen β-glucosidases in filamentous fungi Aspergillus niger. Two of them belonged to glycoside hydrolase family 1 (GH1) while the rest belonged to genes in family 3 (GH3). These proteins were then named according to the nomenclature as proposed by the International Union of Biochemistry (IUB), starting from the lowest pI and glycoside hydrolase family. Their properties were predicted using various bionformatic tools showing the presence of domains for signal peptide and active sites. Interestingly, one particular domain, PA14 (protective antigen) was present in four of the enzymes, predicted to be involved in carbohydrate binding. A phylogenetic tree grouped the two glycoside hydrolase families with GH1 and GH3 related organisms. This study showed that the various domains present in these β-glucosidases are postulated to be crucial for the survival of this fungus, as supported by other analysis.

  18. Tandem shock waves to enhance genetic transformation of Aspergillus niger.

    Science.gov (United States)

    Loske, Achim M; Fernández, Francisco; Magaña-Ortíz, Denis; Coconi-Linares, Nancy; Ortíz-Vázquez, Elizabeth; Gómez-Lim, Miguel A

    2014-08-01

    Filamentous fungi are used in several industries and in academia to produce antibiotics, metabolites, proteins and pharmaceutical compounds. The development of valuable strains usually requires the insertion of recombinant deoxyribonucleic acid; however, the protocols to transfer DNA to fungal cells are highly inefficient. Recently, underwater shock waves were successfully used to genetically transform filamentous fungi. The purpose of this research was to demonstrate that the efficiency of transformation can be improved significantly by enhancing acoustic cavitation using tandem (dual-pulse) shock waves. Results revealed that tandem pressure pulses, generated at a delay of 300 μs, increased the transformation efficiency of Aspergillus niger up to 84% in comparison with conventional (single-pulse) shock waves. This methodology may also be useful to obtain new strains required in basic research and biotechnology. Copyright © 2014 Elsevier B.V. All rights reserved.

  19. Mapping the polysaccharide degradation potential of Aspergillus niger

    DEFF Research Database (Denmark)

    Andersen, Mikael Rørdam; Giese, Malene; de Vries, Ronald P.

    2012-01-01

    Background: The degradation of plant materials by enzymes is an industry of increasing importance. For sustainable production of second generation biofuels and other products of industrial biotechnology, efficient degradation of non-edible plant polysaccharides such as hemicellulose is required...... of a given fungus for polysaccharide degradation. Results: Through the compilation of information from 203 articles, we have systematized knowledge on the structure and degradation of 16 major types of plant polysaccharides to form a graphical overview. As a case example, we have combined this with a list...... of 188 genes coding for carbohydrate-active enzymes from Aspergillus niger, thus forming an analysis framework, which can be queried. Combination of this information network with gene expression analysis on mono-and polysaccharide substrates has allowed elucidation of concerted gene expression from...

  20. Effect of maltose on glucoamylase formation by Aspergillus niger.

    Science.gov (United States)

    Barton, L L; Georgi, C E; Lineback, D R

    1972-09-01

    Low levels of glucoamylase are produced when Aspergillus niger is grown on sorbitol, but substitution of the latter by glucose, maltose, or starch results in greater formation of glucoamylase as measured by enzymatic activity. Both glucoamylase I and glucoamylase II are formed in a yeast extract medium; however, glucoamylase I appears to be the only form produced when ammonium chloride is the nitrogen source. Maltose or isomaltose (1.4 x 10(-4)m), but no other disaccharides or monosaccharides, dextrins, dextrans, or starches, stimulated glucoamylase formation when added to mycelia pregrown on sorbitol-ammonium salts. The induction of glucoamylase by maltose was independent of sulfate concentration but showed a dependency on low pH and the absence of utilizable carbon sources.

  1. Nanosulfur: A Potent Fungicide Against Food Pathogen, Aspergillus niger

    Science.gov (United States)

    Choudhury, Samrat Roy; Nair, Kishore K.; Kumar, Rajesh; Gogoi, Robin; Srivastava, Chitra; Gopal, Madhuban; Subhramanyam, B. S.; devakumar, C.; Goswami, Arunava

    2010-10-01

    Elemental sulfur (S0), man's oldest eco-friendly fungicide for curing fungal infections in plants and animals, is registered in India as a non-systemic and contact fungicide. However due to its high volume requirement, Indian agrochemical industry and farmers could not effectively use this product till date. We hypothesize that intelligent nanoscience applications might increase the visibility of nanosulfur in Indian agriculture as a potent and eco-safe fungicide. Sulfur nanoparticles (NPs) were synthesized bottom-up via a liquid synthesis method with average particle size in the range of 50-80 nm and the shapes of the NPs were spherical. A comparative study of elemental and nano-sulfur produced has been tested against facultative fungal food pathogen, Aspergillus niger. Results showed that nanosulfur is more efficacious than its elemental form.

  2. Enhanced citrate production through gene insertion in Aspergillus niger

    DEFF Research Database (Denmark)

    Jongh, Wian de; Nielsen, Jens

    2007-01-01

    The effect of inserting genes involved in the reductive branch of the tricarboxylic acid (TCA) cycle on citrate production by Aspergillus niger was evaluated. Several different genes were inserted individually and in combination, i.e. malate dehydrogenase (mdh2) from Saccharomyces cerevisiae, two...... truncated, cytosolic targeted, fumarases (Fum1s and FumRs) from S. cerevisiae and Rhizopus oryzae, respectively, and the cytosolic soluble fumarate reductase (Frds1) from S. cerevisiae. Overexpression of these genes in their native strain backgrounds has been reported to lead to alterations...... in the intracellular cytosolic dicarboxylate concentrations. It was found that all the transformant strains had enhanced yield and productivities of citrate compared with the wild-type strain. The transformants also had the ability to produce citrate in trace-manganese-contaminated medium, where the wild type...

  3. Screening a strain of Aspergillus niger and optimization of fermentation conditions for degradation of aflatoxin B

    National Research Council Canada - National Science Library

    Zhang, Wei; Xue, Beibei; Li, Mengmeng; Mu, Yang; Chen, Zhihui; Li, Jianping; Shan, Anshan

    2014-01-01

    ...) was able to degrade aflatoxin B₁ after screening. ND-1 isolate, identified as a strain of Aspergillus niger using phylogenetic analysis on the basis of 18S rDNA, could remove 26.3% of aflatoxin B₁...

  4. Rewiring a secondary metabolite pathway towards itaconic acid production in Aspergillus niger

    NARCIS (Netherlands)

    Hossain, A.H.; Li, A.; Brickwedde, A.; Wilms, L.; Caspers, M.; Overkamp, K.; Punt, P.J.

    2016-01-01

    Background: The industrially relevant filamentous fungus Aspergillus niger is widely used in industry for its secretion capabilities of enzymes and organic acids. Biotechnologically produced organic acids promise to be an attractive alternative for the chemical industry to replace petrochemicals.

  5. Control of Aspergillus niger with garlic, onion and leek extracts | Irkin ...

    African Journals Online (AJOL)

    Allium cepa L.) and leek (Allium porrum L.) were investigated against Aspergillus niger. Minimal inhibitory concentrations (MIC) and minimal fungicidal concentrations (MFC) of aqueous, ethyl alcohol and acetone extracts were determined by ...

  6. Enzymatic Synthesis of Hydroxycinnamic Acid Glycerol Esters Using Type A Feruloyl Esterase from Aspergillus niger

    National Research Council Canada - National Science Library

    TSUCHIYAMA, Moriyasu; SAKAMOTO, Tatsuji; TANIMORI, Shinji; MURATA, Shuichi; KAWASAKI, Haruhiko

    2007-01-01

    We found that hydroxycinnamic acid (HA) glycerol esters such as 1-sinapoyl glycerol and 1-p-coumaroyl glycerol can be synthesized through a direct esterification reaction using a type A feruloyl esterase from Aspergillus niger...

  7. Improving the extraction conditions of endoglucanase produced by Aspergillus niger under solid-state fermentation

    National Research Council Canada - National Science Library

    Pirota, R. D. P. B; Miotto, L. S; Delabona, P. S; Farinas, C. S

    2013-01-01

    ...) on the recovery of endoglucanases produced by Aspergillus niger cultivated under SSF. Statistical analysis revealed that only the solid to liquid ratio had a significant influence on endoglucanase extraction...

  8. Bioleaching of spent fluid catalytic cracking catalyst using Aspergillus niger.

    Science.gov (United States)

    Aung, Khin Moh Moh; Ting, Yen-Peng

    2005-03-16

    The use of the fungus Aspergillus niger for the bioleaching of heavy metals from spent catalyst was investigated, with fluid catalytic cracking (FCC) catalyst as a model. Bioleaching was examined in batch cultures with the spent catalysts at various pulp densities (1-12%). Chemical leaching was also performed using mineral acids (sulphuric and nitric acids) and organic acids (citric, oxalic and gluconic acids), as well as a mixture of organic acids at the same concentrations as that biogenically produced. It was shown that bioleaching realised higher metal extraction than chemical leaching, with A. niger mobilizing Ni (9%), Fe (23%), Al (30%), V (36%) and Sb (64%) at 1% pulp density. Extraction efficiency generally decreased with increased pulp density. Compared with abiotic controls, bioleaching gave rise to higher metal extractions than leaching using fresh medium and cell-free spent medium. pH decreased during bioleaching, but remained relatively constant in both leaching using fresh medium and cell-free spent medium, thus indicating that the fungus played a role in effecting metal extraction from the spent catalyst.

  9. Mapping the polysaccharide degradation potential of Aspergillus niger

    Science.gov (United States)

    2012-01-01

    Background The degradation of plant materials by enzymes is an industry of increasing importance. For sustainable production of second generation biofuels and other products of industrial biotechnology, efficient degradation of non-edible plant polysaccharides such as hemicellulose is required. For each type of hemicellulose, a complex mixture of enzymes is required for complete conversion to fermentable monosaccharides. In plant-biomass degrading fungi, these enzymes are regulated and released by complex regulatory structures. In this study, we present a methodology for evaluating the potential of a given fungus for polysaccharide degradation. Results Through the compilation of information from 203 articles, we have systematized knowledge on the structure and degradation of 16 major types of plant polysaccharides to form a graphical overview. As a case example, we have combined this with a list of 188 genes coding for carbohydrate-active enzymes from Aspergillus niger, thus forming an analysis framework, which can be queried. Combination of this information network with gene expression analysis on mono- and polysaccharide substrates has allowed elucidation of concerted gene expression from this organism. One such example is the identification of a full set of extracellular polysaccharide-acting genes for the degradation of oat spelt xylan. Conclusions The mapping of plant polysaccharide structures along with the corresponding enzymatic activities is a powerful framework for expression analysis of carbohydrate-active enzymes. Applying this network-based approach, we provide the first genome-scale characterization of all genes coding for carbohydrate-active enzymes identified in A. niger. PMID:22799883

  10. Solubilisation of some naturally occurring metal-bearing minerals, limescale and lead phosphate by Aspergillus niger.

    Science.gov (United States)

    Sayer, J A; Kierans, M; Gadd, G M

    1997-09-01

    The ability of the soil fungus Aspergillus niger to tolerate and solubilise seven naturally occurring metal-bearing minerals, limescale and lead phosphate was investigated. A. niger was able to solubilise four of the test insoluble compounds when incorporated into solid medium: cuprite (CuO2), galena (PbS), rhodochrosite (Mn(CO3)x) and limescale (CaCO3). A. niger was able to grow on all concentrations of all the test compounds, whether solubilisation occurred or not, with no reduction in growth rate from the control. In some cases, stimulation of growth occurred, most marked with the phosphate-containing mineral, apatite. Precipitation of insoluble copper and manganese oxalate crystals under colonies growing on agar amended with cuprite and rhodochrosite was observed after 1-2 days growth at 25 degrees C. This process of oxalate formation represents a reduction in bioavailability of toxic cations, and could represent an important means of toxic metal immobilisation of physiological and environmental significance.

  11. Enhanced itaconic acid production in Aspergillus niger using genetic modification and medium optimization

    OpenAIRE

    Li An; Pfelzer Nina; Zuijderwijk Robbert; Punt Peter

    2012-01-01

    Abstract Background Aspergillus niger was selected as a host for producing itaconic acid due to its versatile and tolerant character in various growth environments, and its extremely high capacity of accumulating the precursor of itaconic acid: citric acid. Expressing the CAD gene from Aspergillus terreus opened the metabolic pathway towards itaconic acid in A. niger. In order to increase the production level, we continued by modifying its genome and optimizing cultivation media. Results Base...

  12. Fatal invasive aspergillosis caused by Aspergillus niger after bilateral lung transplantation

    Directory of Open Access Journals (Sweden)

    Enora Atchade

    2017-09-01

    Full Text Available Aspergillus niger is usually considered to be a low virulence fungus, not commonly reported to cause invasive infections. Invasive pulmonary aspergillosis due to Aspergillus niger was diagnosed in a 43-year-old woman following bilateral lung transplantation. Intravenous voriconazole failed to control progression of the disease. Despite salvage therapy with a combination of voriconazole and caspofungin for 23 days, the patient developed massive hemoptysis leading to death. The authors report the clinical features and treatment of this case.

  13. Elucidation of the biosynthesis of meroterpenoid yanuthone D in Aspergillus Niger

    DEFF Research Database (Denmark)

    Holm, Dorte Koefoed; Petersen, Lene Maj; Klitgaard, Andreas

    2012-01-01

    We have elucidated the mode of biosynthesis of the meroterpenoid compound Yanuthone D in Aspergillus niger. We have successfully deleted all cluster genes, and identified a number of intermediates. Structures of the intermediates were solved using a combined approach comprising classical 1D- and ......-NMR and tandem mass spectrometry (MS/MS). In this study we have confirmed that Yanuthone D is of meroterpenoid origin, and we have identified an unexpected precursor, which has not before been reported for Aspergillus niger....

  14. Genome sequencing and analysis of the versatile cell factory Aspergillus niger CBS 513.88

    DEFF Research Database (Denmark)

    Pel, Herman J.; de Winde, Johannes H.; Archer, David B.

    2007-01-01

    The filamentous fungus Aspergillus niger is widely exploited by the fermentation industry for the production of enzymes and organic acids, particularly citric acid. We sequenced the 33.9-megabase genome of A. niger CBS 513.88, the ancestor of currently used enzyme production strains. A high level...

  15. Database mining and transcriptional analysis of genes encoding inulin-modifying enzymes of Aspergillus niger

    NARCIS (Netherlands)

    Yuan, X.L.; Goosen, C.; Kools, H.J.; Maarel, van der M.J.; Hondel, van den C.A.M.J.J.; Dijkhuizen, L.; Ram, A.F.

    2006-01-01

    As a soil fungus, Aspergillus niger can metabolize a wide variety of carbon sources, employing sets of enzymes able to degrade plant-derived polysaccharides. In this study the genome sequence of A. niger strain CBS 513.88 was surveyed, to analyse the gene/enzyme network involved in utilization of

  16. Quantitative iTRAQ secretome analysis of aspergillus niger reveals novel hydrolytic enzymes

    NARCIS (Netherlands)

    Adav, S.S.; Li, A.A.; Manavalan, A.; Punt, P.; Sze, S.K.

    2010-01-01

    The natural lifestyle of Aspergillus niger made them more effective secretors of hydrolytic proteins and becomes critical when this species were exploited as hosts for the commercial secretion of heterologous proteins. The protein secretion profile of A. niger and its mutant at different pH was

  17. Identification and characterization of starch and inulin modifying network of Aspergillus niger by functional genomics

    NARCIS (Netherlands)

    Yuan, Xiao-Lian

    2008-01-01

    Aspergillus niger produces a wide variety of carbohydrate hydrolytic enzymes which have potential applications in the baking, starch, textile, food and feed industries. The goal of this thesis is to unravel the molecular mechanisms of starch and inulin modifying network of A. niger, in order to

  18. Characterization of oxylipins and dioxygenase genes in the asexual fungus Aspergillus niger

    NARCIS (Netherlands)

    Wadman, Mayken W; de Vries, Ronald P; Kalkhove, Stefanie I C; Veldink, Gerrit A; Vliegenthart, Johannes F G

    2009-01-01

    BACKGROUND: Aspergillus niger is an ascomycetous fungus that is known to reproduce through asexual spores, only. Interestingly, recent genome analysis of A. niger has revealed the presence of a full complement of functional genes related to sexual reproduction 1. An example of such genes are the

  19. Assessment of the pectinolytic network of Aspergillus niger by functional genomics : insights from the transcriptome

    NARCIS (Netherlands)

    Martens-Uzunova, E.S.

    2008-01-01

    More than a century ago, in 1889, A. Fernbach presented a detailed report about the invertase of Aspergillus niger in the third edition of “Annales De L'institut Pasteur”. Since then, many of the enzymes secreted by A. niger have found a broad range of applications, and today they are produced on an

  20. Genome sequencing and analysis of the versatile cell factory Aspergillus niger CBS 513.88

    NARCIS (Netherlands)

    Pel, Herman J.; de Winde, Johannes H.; Archer, David B.; Dyer, Paul S.; Hofmann, Gerald; Schaap, Peter J.; Turner, Geoffrey; Albang, Richard; Albermann, Kaj; Andersen, Mikael R.; Bendtsen, Jannick D.; Benen, Jacques A. E.; van den Berg, Marco; Breestraat, Stefaan; Caddick, Mark X.; Contreras, Roland; Cornell, Michael; Coutinho, Pedro M.; Danchin, Etienne G. J.; Debets, Alfons J. M.; Dekker, Peter; van Dijck, Piet W. M.; van Dijk, Alard; Dijkhuizen, Lubbert; Driessen, Arnold J. M.; d'Enfert, Christophe; Geysens, Steven; Groot, Gert S. P.; de Groot, Piet W. J.; Guillemette, Thomas; Henrissat, Bernard; Herweijer, Marga; van den Hombergh, Johannes P. T. W.; van den Hondel, Cees A. M. J. J.; van der Heijden, Rene T. J. M.; van der Kaaij, Rachel M.; Klis, Frans M.; Kools, Harrie J.; Kubicek, Christian P.; van Kuyk, Patricia A.; Lauber, Juergen; Lu, Xin; van der Maarel, Marc J. E. C.; Meulenberg, Rogier; Menke, Hildegard; Mortimer, Martin A.; Nielsen, Jens; Oliver, Stephen G.; Olsthoorn, Maurien; Pal, Karoly; van Peij, Noel N. M. E.; Ram, Arthur F. J.; Rinas, Ursula; Roubos, Johannes A.; Sagt, Cees M. J.; Schmoll, Monika; Sun, Jibin; Ussery, David; Varga, Janos; Vervecken, Wouter; de Vondervoort, Peter J. J. van; Wedler, Holger; Wosten, Han A. B.; Zeng, An-Ping; van Ooyen, Albert J. J.; Visser, Jaap; Stam, Hein; Enfert, Christophe d’; Lauber, Jürgen; Goosen, Coenie; de Vries, Ronald P.

    The filamentous fungus Aspergillus niger is widely exploited by the fermentation industry for the production of enzymes and organic acids, particularly citric acid. We sequenced the 33.9-megabase genome of A. niger CBS 513.88, the ancestor of currently used enzyme production strains. A high level of

  1. Genome sequencing and analysis of the versatile cell factory Aspergillus niger CBS 513.88

    NARCIS (Netherlands)

    Pel, H.J.; Winde, J.H. de; Archer, D.B.; Dyer, P.S.; Hofmann, G.; Schaap, P.J.; Turner, G.; Vries, R.P. de; Albang, R.; Albermann, K.; Andersen, M.R.; Bendtsen, J.D.; Benen, J.A.; Berg, M. van den; Breestraat, S.; Caddick, M.X.; Contreras, R.; Cornell, M.; Coutinho, P.M.; Danchin, E.G.; Debets, A.J.; Dekker, P.; Dijck, P.W. van; Dijk, A. van; Dijkhuizen, L.; Driessen, A.J.; D'Enfert, C.; Geysens, S.; Goosen, C.; Groot, G.S.; Groot, P.W. de; Guillemette, T.; Henrissat, B.; Herweijer, M.; Hombergh, J.P. van den; Hondel, C.A. van den; Heijden, R.T.J.M. van der; Kaaij, R.M. van der; Klis, F.M.; Kools, H.J.; Kubicek, C.P.; Kuyk, P.A. van; Lauber, J.; Lu, X.; Maarel, M.J. van der; Meulenberg, R.; Menke, H.; Mortimer, M.A.; Nielsen, J.; Oliver, S.G.; Olsthoorn, M.; Pal, K.; Peij, N.N. van; Ram, A.F.; Rinas, U.; Roubos, J.A.; Sagt, C.M.; Schmoll, M.; Sun, J.; Ussery, D.; Varga, J.; Vervecken, W.; Vondervoort, P.J. van de; Wedler, H.; Wosten, H.A.; Zeng, A.P.; Ooyen, A.J. van; Visser, J.C.; Stam, H.

    2007-01-01

    The filamentous fungus Aspergillus niger is widely exploited by the fermentation industry for the production of enzymes and organic acids, particularly citric acid. We sequenced the 33.9-megabase genome of A. niger CBS 513.88, the ancestor of currently used enzyme production strains. A high level of

  2. Dual transcriptional profiling of a bacterial/fungal confrontation: Collimonas fungivorans versus Aspergillus niger.

    Science.gov (United States)

    Mela, Francesca; Fritsche, Kathrin; de Boer, Wietse; van Veen, Johannes A; de Graaff, Leo H; van den Berg, Marlies; Leveau, Johan H J

    2011-09-01

    Interactions between bacteria and fungi cover a wide range of incentives, mechanisms and outcomes. The genus Collimonas consists of soil bacteria that are known for their antifungal activity and ability to grow at the expense of living fungi. In non-contact confrontation assays with the fungus Aspergillus niger, Collimonas fungivorans showed accumulation of biomass concomitant with inhibition of hyphal spread. Through microarray analysis of bacterial and fungal mRNA from the confrontation arena, we gained new insights into the mechanisms underlying the fungistatic effect and mycophagous phenotype of collimonads. Collimonas responded to the fungus by activating genes for the utilization of fungal-derived compounds and for production of a putative antifungal compound. In A. niger, differentially expressed genes included those involved in lipid and cell wall metabolism and cell defense, which correlated well with the hyphal deformations that were observed microscopically. Transcriptional profiles revealed distress in both partners: downregulation of ribosomal proteins and upregulation of mobile genetic elements in the bacteria and expression of endoplasmic reticulum stress and conidia-related genes in the fungus. Both partners experienced nitrogen shortage in each other's presence. Overall, our results indicate that the Collimonas/Aspergillus interaction is a complex interplay between trophism, antibiosis and competition for nutrients.

  3. GalX regulates the d-galactose oxido-reductive pathway in Aspergillus niger

    NARCIS (Netherlands)

    Gruben, B.S.; Zhou, M.; de Vries, R.P.

    2012-01-01

    Galactose catabolism in Aspergillus nidulans is regulated by at least two regulators, GalR and GalX. In Aspergillus niger only GalX is present, and its role in d-galactose catabolism in this fungus was investigated. Phenotypic and gene expression analysis of a wild type and a galX disruptant

  4. Formation of Sclerotia and Production of Indoloterpenes by Aspergillus niger and Other Species in Section Nigri

    DEFF Research Database (Denmark)

    Frisvad, Jens Christian; Petersen, Lene Maj; Lyhne, Ellen Kirstine

    2014-01-01

    Several species in Aspergillus section Nigri have been reported to produce sclerotia on well-known growth media, such as Czapek yeast autolysate (CYA) agar, with sclerotia considered to be an important prerequisite for sexual development. However Aspergillus niger sensu stricto has not been repor...

  5. Biocatalytic potential of laccase-like multicopper oxidases from Aspergillus niger

    NARCIS (Netherlands)

    Tamayo Ramos, J.A.; Berkel, van W.J.H.; Graaff, de L.H.

    2012-01-01

    BACKGROUND: Laccase-like multicopper oxidases have been reported in several Aspergillus species but they remain uncharacterized. The biocatalytic potential of the Aspergillus niger fungal pigment multicopper oxidases McoA and McoB and ascomycete laccase McoG was investigated. RESULTS: The

  6. Enhanced itaconic acid production in Aspergillus niger using genetic modification and medium optimization

    NARCIS (Netherlands)

    Li, A.; Pfelzer, N.; Zuijderwijk, R.; Punt, P.J.

    2012-01-01

    Background: Aspergillus niger was selected as a host for producing itaconic acid due to its versatile and tolerant character in various growth environments, and its extremely high capacity of accumulating the precursor of itaconic acid: citric acid. Expressing the CAD gene from Aspergillus terreus

  7. Post-genomic insights into the plant polysaccharide degradation potential of Aspergillus nidulans and comparison to Aspergillus niger and Aspergillus oryzae

    DEFF Research Database (Denmark)

    Coutinho, Pedro M.; Andersen, Mikael Rørdam; Kolenova, Katarina

    2009-01-01

    The plant polysaccharide degradative potential of Aspergillus nidulans was analysed in detail and compared to that of Aspergillus niger and Aspergillus oryzae using a combination of bioinformatics, physiology and transcriptomics. Manual verification indicated that 28.4% of the A. nidulans ORFs...... on polysaccharides. Growth differences were observed between the Aspergilli and Podospora anserina, which has a more different genomic potential for polysaccharide degradation, suggesting that large genomic differences are required to cause growth differences oil polysaccharides, Differences were also detected...

  8. Identification and cloning of a second phytase gene (phyB) from Aspergillus niger (ficuum).

    Science.gov (United States)

    Ehrlich, K C; Montalbano, B G; Mullaney, E J; Dischinger, H C; Ullah, A H

    1993-08-31

    An Aspergillus niger (ficuum) genomic DNA lambda EMBL3 library was probed with a 354-bp DNA fragment obtained by polymerase chain reaction of A. niger DNA with oligonucleotides based on partial amino acid sequence of a pH 2.5 optimum acid phosphatase. A clone containing a 1605 bp segment (phyB) encoding the 479 amino acid enzyme was isolated and found to contain four exons. Global alignment revealed 23.5% homology to Aspergillus niger phytase (PhyA); four regions of extensive homology were identified. Some of these regions may contain catalytic sites for phosphatase function.

  9. Fumonisin B2 production by Aspergillus niger in Thai coffee beans

    DEFF Research Database (Denmark)

    Noonim, P.; Mahakarnchanaku, W.; Nielsen, Kristian Fog

    2009-01-01

    During 2006 and 2007, a total of 64 Thai dried coffee bean samples (Coffea arabica) from two growing sites in Chiangmai Province and 32 Thai dried coffee bean samples (Coffea canephora) from two growing sites in Chumporn Province, Thailand, were collected and assessed for fumonisin contamination...... by black Aspergilli. No Fusarium species known to produce fumonisin were detected, but black Aspergilli had high incidences on both Arabica and Robusta Thai coffee beans. Liquid chromatography (LC) with high-resolution mass spectrometric (HRMS) detection showed that 67% of Aspergillus niger isolates from...... coffee beans were capable of producing fumonisins B2 (FB2) and B4 when grown on Czapek Yeast Agar with 5% NaCl. Small amounts (1-9.7 ng g-1) of FB2 were detected in seven of 12 selected coffee samples after ion-exchange purification and LC-MS/MS detection. Two samples also contained FB4...

  10. Mapping the polysaccharide degradation potential of Aspergillus niger

    Directory of Open Access Journals (Sweden)

    Andersen Mikael R

    2012-07-01

    Full Text Available Abstract Background The degradation of plant materials by enzymes is an industry of increasing importance. For sustainable production of second generation biofuels and other products of industrial biotechnology, efficient degradation of non-edible plant polysaccharides such as hemicellulose is required. For each type of hemicellulose, a complex mixture of enzymes is required for complete conversion to fermentable monosaccharides. In plant-biomass degrading fungi, these enzymes are regulated and released by complex regulatory structures. In this study, we present a methodology for evaluating the potential of a given fungus for polysaccharide degradation. Results Through the compilation of information from 203 articles, we have systematized knowledge on the structure and degradation of 16 major types of plant polysaccharides to form a graphical overview. As a case example, we have combined this with a list of 188 genes coding for carbohydrate-active enzymes from Aspergillus niger, thus forming an analysis framework, which can be queried. Combination of this information network with gene expression analysis on mono- and polysaccharide substrates has allowed elucidation of concerted gene expression from this organism. One such example is the identification of a full set of extracellular polysaccharide-acting genes for the degradation of oat spelt xylan. Conclusions The mapping of plant polysaccharide structures along with the corresponding enzymatic activities is a powerful framework for expression analysis of carbohydrate-active enzymes. Applying this network-based approach, we provide the first genome-scale characterization of all genes coding for carbohydrate-active enzymes identified in A. niger.

  11. Production and Characterization of Glucoamylase by Aspergillus niger

    Directory of Open Access Journals (Sweden)

    Soumik Banerjee

    2017-01-01

    Full Text Available Background and Objective: Glucoamylase is a potent starch degrading enzyme whose cheap production has been an area of research. Its production by Aspergillus niger in solid-state fermentation was studied using dried garden pea peel as a substrate, which enormously reduced the production cost. The current study intended to produce glucoamylase by a cost-effective strategy and exhaustively characterize the enzyme.Material and Methods: Garden pea peel was used as a substrate in solid state fermentation by Aspergillus niger for the production of glucoamylase under process parameters. Response surface methodology, a statistical tool for optimization, was applied to setup the experimental design for glucoamylase production. Characterization studies of the enzyme were carried out with temperature, pH, metal salts and elemental composition analysis.Results and Conclusion: The process parameters were temperature, amount of substrate and time of fermentation. Glucoamylase production was highest in the pH range of 5.4-6.2, was stable at pH 3.8, and maintained its maximum activity even at 70°C for 30 min. It showed higher catalytic efficiency when incubated with metal ions Fe2+, Cu2+, Mg2+, and Pb2+. Km and Vmax for glucoamylase were 0.387 mg of soluble starch ml-1 and 35.03 U μl-1 min-1, respectively. Glycogen was also used as a substrate, which gave an increased Km by 2.585, whose KI was found to be 0.631. Energy-dispersive X-ray spectroscopy was performed for obtaining composition of the pea peel. C, N, and O were found to be 12.53%, 29.9%, and 55.27% by atomic weights, respectively. Cost- and time-effective production of glucoamylase was achieved by utilizing dried garden pea peel (a vegetable residue powder as the substrate for production. Its high stability ensures efficient utilization under industrial conditions. This work provides a very good platform for the enzyme immobilization studies and scale up production in future.Conflict of interest

  12. A novel fungal fruiting structure formed by Aspergillus niger and Aspergillus carbonarius in grape berries.

    Science.gov (United States)

    Pisani, Cristina; Nguyen, Trang Thoaivan; Gubler, Walter Douglas

    2015-09-01

    Sour rot, is a pre-harvest disease that affects many grape varieties. Sour rot symptoms include initial berry cracking and breakdown of berry tissue. This is a disease complex with many filamentous fungi and bacteria involved, but is usually initiated by Aspergillus niger or Aspergillus carbonarius. Usually, by the time one sees the rot there are many other organisms involved and it is difficult to attribute the disease to one species. In this study two species of Aspergillus were shown to produce a previously unknown fruiting structure in infected berries. The nodulous morphology, bearing conidia, suggests them to be an 'everted polymorphic stroma'. This structure forms freely inside the berry pulp and assumes multiple shapes and sizes, sometimes sclerotium-like in form. It is composed of a mass of vegetative hyphae with or without tissue of the host containing spores or fruiting bodies bearing spores. Artificially inoculated berries placed in soil in winter showed the possible overwintering function of the fruiting body. Inoculated berry clusters on standing vines produced fruiting structures within 21 d post inoculation when wounds were made at veraison or after (July-September). Histological studies confirmed that the fruiting structure was indeed fungal tissue. Copyright © 2015 The British Mycological Society. Published by Elsevier Ltd. All rights reserved.

  13. Phytase Production by Aspergillus niger CFR 335 and Aspergillus ficuum SGA 01 through Submerged and Solid-State Fermentation

    OpenAIRE

    Shivanna, Gunashree B.; Govindarajulu Venkateswaran

    2014-01-01

    Fermentation is one of the industrially important processes for the development of microbial metabolites that has immense applications in various fields. This has prompted to employ fermentation as a major technique in the production of phytase from microbial source. In this study, a comparison was made between submerged (SmF) and solid-state fermentations (SSF) for the production of phytase from Aspergillus niger CFR 335 and Aspergillus ficuum SGA 01. It was found that both the fungi were ca...

  14. Screening a strain of Aspergillus niger and optimization of fermentation conditions for degradation of aflatoxin B₁.

    Science.gov (United States)

    Zhang, Wei; Xue, Beibei; Li, Mengmeng; Mu, Yang; Chen, Zhihui; Li, Jianping; Shan, Anshan

    2014-11-13

    Aflatoxin B₁, a type of highly toxic mycotoxin produced by some species belonging to the Aspergillus genus, such as Aspergillus flavus and Aspergillus parasiticus, is widely distributed in feed matrices. Here, coumarin was used as the sole carbon source to screen microorganism strains that were isolated from types of feed ingredients. Only one isolate (ND-1) was able to degrade aflatoxin B₁ after screening. ND-1 isolate, identified as a strain of Aspergillus niger using phylogenetic analysis on the basis of 18S rDNA, could remove 26.3% of aflatoxin B₁ after 48 h of fermentation in nutrient broth (NB). Optimization of fermentation conditions for aflatoxin B₁ degradation by selected Aspergillus niger was also performed. These results showed that 58.2% of aflatoxin B₁ was degraded after 24 h of culture under the optimal fermentation conditions. The aflatoxin B₁ degradation activity of Aspergillus niger supernatant was significantly stronger than cells and cell extracts. Furthermore, effects of temperature, heat treatment, pH, and metal ions on aflatoxin B₁ degradation by the supernatant were examined. Results indicated that aflatoxin B₁ degradation of Aspergillus niger is enzymatic and this process occurs in the extracellular environment.

  15. Physiological optimization of secreted protein production by Aspergillus niger.

    Science.gov (United States)

    MacKenzie, D A; Gendron, L C; Jeenes, D J; Archer, D B

    1994-04-01

    Physiological factors affecting hen eggwhite lysozyme and native glucoamylase production by Aspergillus niger have been examined in batch culture. Expression of the genes encoding both proteins was controlled by the glucoamylase promoter. In standard expression medium (ACMS/N/P), secreted lysozyme yields were found to be maximal at 20-25 degrees C (8-10 mg l-1) and markedly reduced at 30-37 degrees C (3-5 mg l-1). Production of lysozyme exhibited similar induction or repression profiles to that of endogenous glucoamylase such that secreted lysozyme yields could be ordered with respect to growth on the following carbon sources: soluble starch > maltose > glucose > xylose. Significantly higher yields of up to 30-60 mg l-1 were obtained in a richer medium containing soya milk, although in contrast to growth in ACMS/N/P, the highest levels of secreted lysozyme were achieved at 37 degrees C. This improvement is attributed partly to an increase in culture biomass concentration and to a reduction in medium acidification. Growth in this medium produced a markedly different pellet morphology.

  16. Catalytic Properties of Lipase Extracts from Aspergillus niger

    Directory of Open Access Journals (Sweden)

    Cintia M. Romero

    2006-01-01

    Full Text Available Screening of lipolytic strains using Rhodamine-B/olive oil plate technique allowed the selection of Aspergillus niger MYA 135. Lipase production in submerged culture containing 2 % olive oil was enhanced by more than 50 % compared to basal cultural conditions. Optimal catalytic conditions for olive oil-induced lipase were pH=6.5 and 30–35 °C. These values were shifted to the acid region (4.0–6.5 and 35–37 °C when lipase extract was produced under basal conditions. Slight changes of the residual lipase activity against the pH were found. However, preincubation at either 37 or 40 °C caused an increase in the olive oil-inducible lipolytic activity. On the contrary, lipase residual activity decreases in the 30–55 °C range when it was produced in basal medium. Lipolytic extracts were almost not deactivated in presence of 50 % water-miscible organic solvents. However, water-immiscible aliphatic solvents reduced the lipase activity between 20 and 80 %.

  17. Switching from a unicellular to multicellular organization in an Aspergillus niger hypha.

    Science.gov (United States)

    Bleichrodt, Robert-Jan; Hulsman, Marc; Wösten, Han A B; Reinders, Marcel J T

    2015-03-03

    Pores in fungal septa enable cytoplasmic streaming between hyphae and their compartments. Consequently, the mycelium can be considered unicellular. However, we show here that Woronin bodies close ~50% of the three most apical septa of growing hyphae of Aspergillus niger. The incidence of closure of the 9th and 10th septa was even ≥94%. Intercompartmental streaming of photoactivatable green fluorescent protein (PA-GFP) was not observed when the septa were closed, but open septa acted as a barrier, reducing the mobility rate of PA-GFP ~500 times. This mobility rate decreased with increasing septal age and under stress conditions, likely reflecting a regulatory mechanism affecting septal pore diameter. Modeling revealed that such regulation offers effective control of compound concentration between compartments. Modeling also showed that the incidence of septal closure in A. niger had an even stronger impact on cytoplasmic continuity. Cytoplasm of hyphal compartments was shown not to be in physical contact when separated by more than 4 septa. Together, data show that apical compartments of growing hyphae behave unicellularly, while older compartments have a multicellular organization. The hyphae of higher fungi are compartmentalized by porous septa that enable cytosolic streaming. Therefore, it is believed that the mycelium shares cytoplasm. However, it is shown here that the septa of Aspergillus niger are always closed in the oldest part of the hyphae, and therefore, these compartments are physically isolated from each other. In contrast, only part of the septa is closed in the youngest part of the hyphae. Still, compartments in this hyphal part are physically isolated when separated by more than 4 septa. Even open septa act as a barrier for cytoplasmic mixing. The mobility rate through such septa reduces with increasing septal age and under stress conditions. Modeling shows that the septal pore width is set such that its regulation offers maximal control of

  18. Optimization of citric acid production by Aspergillus niger

    Directory of Open Access Journals (Sweden)

    H.R Samadlouie

    2015-11-01

    Full Text Available Among the various fungal strains screened for citric acid production, Aspergillus niger is known to produce considerable amounts of citric acid and other organic acids when cultivated in submerge fermentation. In this study, optimization of the medium components was carried out using "one-factor-at-a-time" and response surface methods (RSM. One-factor-at-a-time indicated that the amount of citric acid production was increased along with the increasing of agitation speed (from 150 to 200 rpm, raising the incubation temperature (from 17 to 32 and decreasing pH value (to 2. Moreover, in comparison with the other groups, citric acid production in the treatment containing soy bean powder and minerals revealed a significant increase (up to 25 g/l. The results showed that application of ultrasonic wave during the growth phase could remarkably enhance the production of citric acid. Based on the results of one-factor-at-a-time, sucrose and soy bean powder were the selected additives to test their effect on citric acid production using RSM. The two variables were identified to have significant effects on citric acid production and the maximum citric acid production of 58 g/l was resulted from the combination of 230.87 g/l sucrose and 200.81 g/l soy bean powder. In this study, the significant increase in the production of citric acid in the optimal conditions indicated the using of appropriate statistical methods and also the correct levels of selected variables.

  19. Gen- und verfahrenstechnische Ansätze zur Optimierung von Aspergillus niger als Expressionsplattform

    OpenAIRE

    Wanka, Franziska

    2016-01-01

    Aspergillus niger has the ability to synthesise large amounts of acids and proteins, and to secrete these into the environment. This particular property is used in biotechnology to produce platform chemicals and industrial enzymes. Despite impressive metabolic activities, there is still a need for research to optimise product yields of A. niger using genetic and engineering approaches. This work therefore focuses on using genetic approaches to adapt synthetic gene switches, such as tetracy...

  20. Invasive Aspergillus niger complex infections in a Belgian tertiary care hospital.

    Science.gov (United States)

    Vermeulen, E; Maertens, J; Meersseman, P; Saegeman, V; Dupont, L; Lagrou, K

    2014-05-01

    The incidence of invasive infections caused by the Aspergillus niger species complex was 0.043 cases/10 000 patient-days in a Belgian university hospital (2005-2011). Molecular typing was performed on six available A. niger complex isolates involved in invasive disease from 2010 to 2011, revealing A. tubingensis, which has higher triazole minimal inhibitory concentrations, in five out of six cases. © 2013 The Authors Clinical Microbiology and Infection © 2013 European Society of Clinical Microbiology and Infectious Diseases.

  1. Citric Acid Fermentation by Aspergillus niger on Low Sugar Concentrations and Cotton Waste

    OpenAIRE

    Kiel, Hildegard; Guvrin, Rumia; Henis, Yigal

    1981-01-01

    The possible use of cotton waste as a carbohydrate source of citric acid production by Aspergillus niger was examined. No citric acid was produced when A. niger was grown on cotton waste as a sole carbon source. In two-stage fermentations, however, mycelium obtained from surface cultures in cotton waste medium yielded more citric acid when transferred to sucrose-containing media than when directly inoculated to sucrose-containing media. It is concluded that cotton waste can be used for saving...

  2. The infrared spectral transmittance of Aspergillus niger spore aggregated particle swarm

    Science.gov (United States)

    Zhao, Xinying; Hu, Yihua; Gu, Youlin; Li, Le

    2015-10-01

    Microorganism aggregated particle swarm, which is quite an important composition of complex media environment, can be developed as a new kind of infrared functional materials. Current researches mainly focus on the optical properties of single microorganism particle. As for the swarm, especially the microorganism aggregated particle swarm, a more accurate simulation model should be proposed to calculate its extinction effect. At the same time, certain parameters deserve to be discussed, which helps to better develop the microorganism aggregated particle swarm as a new kind of infrared functional materials. In this paper, take Aspergillus Niger spore as an example. On the one hand, a new calculation model is established. Firstly, the cluster-cluster aggregation (CCA) model is used to simulate the structure of Aspergillus Niger spore aggregated particle. Secondly, the single scattering extinction parameters for Aspergillus Niger spore aggregated particle are calculated by using the discrete dipole approximation (DDA) method. Thirdly, the transmittance of Aspergillus Niger spore aggregated particle swarm is simulated by using Monte Carlo method. On the other hand, based on the model proposed above, what influences can wavelength causes has been studied, including the spectral distribution of scattering intensity of Aspergillus Niger spore aggregated particle and the infrared spectral transmittance of the aggregated particle swarm within the range of 8~14μm incident infrared wavelengths. Numerical results indicate that the scattering intensity of Aspergillus Niger spore aggregated particle reduces with the increase of incident wavelengths at each scattering angle. Scattering energy mainly concentrates on the scattering angle between 0~40°, forward scattering has an obvious effect. In addition, the infrared transmittance of Aspergillus Niger spore aggregated particle swarm goes up with the increase of incident wavelengths. However, some turning points of the trend

  3. Production of Proteolytic Enzymes by a Keratin-Degrading Aspergillus niger

    Science.gov (United States)

    Lopes, Fernanda Cortez; Silva, Lucas André Dedavid e; Tichota, Deise Michele; Daroit, Daniel Joner; Velho, Renata Voltolini; Pereira, Jamile Queiroz; Corrêa, Ana Paula Folmer; Brandelli, Adriano

    2011-01-01

    A fungal isolate with capability to grow in keratinous substrate as only source of carbon and nitrogen was identified as Aspergillus niger using the sequencing of the ITS region of the rDNA. This strain produced a slightly acid keratinase and an acid protease during cultivation in feather meal. The peak of keratinolytic activity occurred in 48 h and the maximum proteolytic activity in 96 h. These enzymes were partly characterized as serine protease and aspartic protease, respectively. The effects of feather meal concentration and initial pH on enzyme production were evaluated using a central composite design combined with response surface methodology. The optimal conditions were determined as pH 5.0 for protease and 7.8 for keratinase and 20 g/L of feather meal, showing that both models were predictive. Production of keratinases by A. niger is a less-exploited field that might represent a novel and promising biotechnological application for this microorganism. PMID:22007293

  4. Production of Proteolytic Enzymes by a Keratin-Degrading Aspergillus niger

    Directory of Open Access Journals (Sweden)

    Fernanda Cortez Lopes

    2011-01-01

    Full Text Available A fungal isolate with capability to grow in keratinous substrate as only source of carbon and nitrogen was identified as Aspergillus niger using the sequencing of the ITS region of the rDNA. This strain produced a slightly acid keratinase and an acid protease during cultivation in feather meal. The peak of keratinolytic activity occurred in 48 h and the maximum proteolytic activity in 96 h. These enzymes were partly characterized as serine protease and aspartic protease, respectively. The effects of feather meal concentration and initial pH on enzyme production were evaluated using a central composite design combined with response surface methodology. The optimal conditions were determined as pH 5.0 for protease and 7.8 for keratinase and 20 g/L of feather meal, showing that both models were predictive. Production of keratinases by A. niger is a less-exploited field that might represent a novel and promising biotechnological application for this microorganism.

  5. ASPERGILLUS NIGER ASPERGILLUS NIGER

    African Journals Online (AJOL)

    User

    Citric acid is present in essentially all plants and many animal tissues and fluids. It is wine, milk, cheese and it is abundant ... behaviour of a process, it is important to formu dynamic models of such processes. These mo upon calibration and ...... intelligence techniques”. International Journal of. Simulation and Modelling, Vol.

  6. ASPERGILLUS NIGER ASPERGILLUS NIGER

    African Journals Online (AJOL)

    User

    models, Monod, Haldane, logistic and hyperbolic f explored. The validity of the models in terms of pred. The validity of the models in terms of pred ...... 26, Number 2, pp. 36-47. [9] Suja M.R.M. and Thyagarajan, T. “Modelling of continuous stirred tank reactor using artificial intelligence techniques”. International Journal of.

  7. ASPERGILLUS NIGER ASPERGILLUS NIGER

    African Journals Online (AJOL)

    eobe

    Zinc (II) ion, Iron (III) ion and methanol on the production of citric acid. Response surface methodology (RSM) was ethanol on the production of citric acid. Response surface methodology (RSM) was used to optimise the effects of these stimulants. The results of analysis of variance (ANOVA) carried out on the model showed ...

  8. Biochemical characterization of three Aspergillus niger β-galactosidases

    Directory of Open Access Journals (Sweden)

    Dandan Niu

    2017-05-01

    Conclusions: Three new β-galactosidases belonging to glycosyl hydrolase family 35 from A. niger F0215 were cloned and biochemically characterized. In addition to the known LacA, A. niger has at least three β-galactosidase family members with remarkably different biochemical properties.

  9. Inhibition of secretion by asexual reproduction in Aspergillus niger

    NARCIS (Netherlands)

    Wang, Fengfeng|info:eu-repo/dai/nl/32304090X

    2014-01-01

    The aim of this Thesis was to study mechanism involved in zonal secretion in colonies of A. niger with emphasis on sporulation inhibited secretion. These mechanisms may be targets to improve A. nigeras a cell factory. I focused in particular on A. niger homologues of fluG and veA that are known

  10. Corn stover-enhanced cellulase production by Aspergillus niger ...

    African Journals Online (AJOL)

    The production of extracellular cellulases by Aspergilus niger NRRL 567 on corn stover was studied in liquid state fermentation. In this study, three cellulases, exoglucanase (EXG), endoglucanase (EG) and β-glucosidase (BGL) were produced by A. niger NRRL 567. The optimal pH, temperature and incubation time for ...

  11. Application of Aspergillus niger SA1 for the enhanced bioremoval of ...

    African Journals Online (AJOL)

    Biological remediation is always envisaged as cost effective and eco-friendly for the treatment of recalcitrant dyes and effluents. Aspergillus niger SA1, a brown rot fungi, isolated from storage pond of textile wastewater, showed a great mineralizing ability for azo dyes, acid red (AR) 151 and orange (Or) II. Decolorization ...

  12. NIGERLYSINTM, HEMOLYSIN PRODUCED BY ASPERGILLUS NIGER, CAUSES LETHALITY OF PRIMARY RAT CORTICAL NEURONAL CELLS IN VITRO

    Science.gov (United States)

    Aspergillus niger produced a proteinaceous hemolysin, nigerlysinTM when incubated on sheep's blood agar at both 23° C and 37°C. Nigerlysin was purified from tryptic soy broth culture filtrate. Purified nigerlysin has a molecular weight of approximately 72 kDa, with an...

  13. Identification of a Transcription Factor Controlling pH-Dependent Organic Acid Response in Aspergillus niger

    DEFF Research Database (Denmark)

    Poulsen, Lars; Andersen, Mikael Rørdam; Lantz, Anna Eliasson

    2012-01-01

    Acid formation in Aspergillus niger is known to be subjected to tight regulation, and the acid production profiles are fine-tuned to respond to the ambient pH. Based on transcriptome data, putative trans-acting pH responding transcription factors were listed and through knock out studies, mutants...

  14. Mode of action of pectin lyase A of Aspergillus niger on differently C6-substituted oligogalacturonides

    NARCIS (Netherlands)

    Alebeek, van G.J.W.M.; Christensen, T.M.I.E.; Schols, H.A.; Mikkelsen, J.D.; Voragen, A.G.J.

    2002-01-01

    A thorough investigation of the mode of action of Aspergillus niger (4M-147) pectin lyase A (PLA) on differently C6-substituted oligogalacturonides is described. PLA appeared to be very specific for fully methyl-esterified oligogalacturonides: removal of the methyl-ester or changing the type of

  15. Assessment of the pectin degrading enzyme network of Aspergillus niger by functional genomics

    NARCIS (Netherlands)

    Martens-Uzunova, E.S.; Schaap, P.J.

    2009-01-01

    The saprobic fungus Aspergillus niger is an efficient producer of a suite of extracellular enzymes involved in carbohydrate modification and degradation. Genome mining has resulted in the prediction of at least 39 genes encoding enzymes involved in the depolymerisation of the backbone of pectin.

  16. The host range of Aspergillus niger and Fusarium oxysporum in the ...

    African Journals Online (AJOL)

    A study was carried out to determine the host range of two fungal organisms, Fusarium oxysporum, the cause of vegetable root rots and Aspergillus niger, the black mold organism, among the members of the family Solanaceae. The test crops were tomato (Lycopersicon esculentum), pepper (Capsicum annuum), potato ...

  17. Mutagenesis and mitotic recombination in Aspergillus niger, an expedition from gene to genome

    NARCIS (Netherlands)

    Vondervoort, Peter Jozef Ida van de

    2007-01-01

    Aspergillus niger is a cosmopolitan fungus and its spores can be found in air and soil worldwide. This saprophyte is used in food biotechnology for the production of proteins, mainly enzymes and for the production of organic acids. In the production of proteins, several problems are encountered such

  18. Characterization of four new antifungal yanuthones from Aspergillus niger

    DEFF Research Database (Denmark)

    Petersen, Lene Maj; Holm, Dorte Koefoed; Knudsen, Peter Boldsen

    2015-01-01

    Four new yanuthone analogs (1–4) were isolated from the filamentous fungus Aspergillus niger. The structures of the new compounds were elucidated on the basis of UHPLC-DAD-HRMS data and one-dimensional and two-dimensional NMR spectroscopy. Labeling studies with 13C8-6-methylsalicylic acid identif...

  19. Disseminated Aspergillosis due to Aspergillus niger in Immunocompetent Patient: A Case Report

    Directory of Open Access Journals (Sweden)

    Ulku Ergene

    2013-01-01

    Full Text Available Invasive aspergillosis is a major cause of morbidity and mortality in immunocompromised patients. Many cases of pulmonary, cutaneous, cerebral, and paranasal sinus aspergillosis in immunocompetent patient were defined in literature but disseminated aspergillosis is very rare. Here we present an immunocompetent case with extrapulmonary disseminated aspergillosis due to Aspergillus niger, totally recovered after effective antifungal treatment with voriconazole.

  20. Aspergillus niger membrane-associated proteome analysis for the identification of glucose transporters

    NARCIS (Netherlands)

    Sloothaak, J.; Odoni, D.I.; Graaff, de L.H.; Martins dos Santos, V.A.P.; Schaap, P.J.; Tamayo Ramos, J.A.

    2015-01-01

    BACKGROUND: The development of biological processes that replace the existing petrochemical-based industry is one of the biggest challenges in biotechnology. Aspergillus niger is one of the main industrial producers of lignocellulolytic enzymes, which are used in the conversion of lignocellulosic

  1. Crystallization and Preliminary Crystallographic Characterization of Endo-polygalacturonase II from Aspergillus niger

    NARCIS (Netherlands)

    Schröter, K.-H.; Arkema, A.; Kester, H.C.M.; Visser, J.; Dijkstra, B.W.

    1994-01-01

    The endo-polygalacturonase II from Aspergillus niger has been crystallized from an ammonium sulfate solution by the hanging drop method. The crystals belong to the monoclinic space group P2(1), with cell dimensions a = 60.6 Angstrom, b = 152.6 Angstrom, c = 74.0 Angstrom and beta = 91.2 degrees with

  2. Study of the glucoamylase promoter in Aspergillus niger using green fluorescent protein

    NARCIS (Netherlands)

    Santerre Henriksen, A.L.; Even, S.; Müller, C.; Punt, P.J.; Hondel, C.A.M.J.J. van den; Nielsen, J.

    1999-01-01

    An Aspergillus niger strain expressing a red-shifted green fluorescent protein (GFP) in the cytoplasm under the control of the glucoamylase promoter (PglaA) was characterized with respect to its physiology and morphology. Although xylose acted as a repressor carbon source during batch cultivations,

  3. Exo-inulinase of Aspergillus niger N402: A hydrolytic enzyme with significant transfructosylating activity

    NARCIS (Netherlands)

    Goosen, C.; Maarel, M.J. E.C. van der; Dijkhuizen, L.

    2008-01-01

    The purified exo-inulinase enzyme of Aspergillus niger N402 (AngInuE; heterologously expressed in Escherichia coli) displayed a sucrose:inulin (S/I) hydrolysis ratio of 2.3, characteristic for a typical exo-inulinase. The enzyme also had significant transfructosylating activity with increasing

  4. The effect of organic nitrogen sources on recombinant glucoamylase production by Aspergillus niger in chemostat culture

    NARCIS (Netherlands)

    Swift, R.J.; Karandikar, A.; Griffen, A.M.; Punt, P.J.; Hondel, C.A.M.J.J. van den; Robson, G.D.; Trinci, A.P.J.; Wiebe, M.G.

    2000-01-01

    Aspergillus niger B1, a recombinant strain carrying 20 extra copies of the native glucoamylase gene, was grown in glucose-limited chemostat cultures supplemented with various organic nitrogen sources (dilution rate 0.12 ± 0.01 h-1, pH 5.4). In cultures supplemented with L-alanine, L-methionine,

  5. Functional analysis of the transcriptional activator XlnR from Aspergillus niger

    NARCIS (Netherlands)

    Hasper, A.A.; Trindade, L.M.; Veen, van der D.; Ooyen, van A.J.J.; Graaff, de L.H.

    2004-01-01

    The transcriptional activator XlnR from Aspergillus niger is a zinc binuclear cluster transcription factor that belongs to the GAL4 superfamily. Several putative structural domains in XInR were predicted using database and protein sequence analysis. Thus far, only the functionality of the N-terminal

  6. Development of tools for quantitative intracellular metabolomics of Aspergillus niger chemostat cultures

    NARCIS (Netherlands)

    Lameiras, F.; Heijnen, J.J.; Van Gulik, W.M.

    2015-01-01

    In view of the high citric acid production capacity of Aspergillus niger, it should be well suited as a cell factory for the production of other relevant acids as succinic, fumaric, itaconic and malic. Quantitative metabolomics is an important omics tool in a synthetic biology approach to develop A.

  7. Production of bio-ethanol from corncobs using Aspergillus niger and ...

    African Journals Online (AJOL)

    Maize is the most abundant cereal grown in Ghana and is accompanied by enormous amount of agrowastes of which corncobs form 30%. This agrowaste which is currently under utilized was used to produce bio-ethanol. Aspergillus niger isolated from soil sampled from Ejura farms was used to hydrolyze the corncobs into ...

  8. The effect of environmental conditions on extracellular protease activity in controlled fermentations of Aspergillus niger

    NARCIS (Netherlands)

    Braaksma, M.; Smilde, A.K.; Werf, M.J. van der; Punt, P.J.

    2009-01-01

    Proteolytic degradation by host proteases is one of the key issues in the application of filamentous fungi for non-fungal protein production. In this study the influence of several environmental factors on the production of extracellular proteases of Aspergillus niger was investigated systematically

  9. Effects of Aspergillus niger (K8) on nutritive value of rice straw ...

    African Journals Online (AJOL)

    The objective of this study was to evaluate the use of solid state fermentation for the improvement of the quality of rice straw as animal feed. Rice straw was fermented using Aspergillus niger (K8) with and without additional nitrogen source (urea). Cellulose, hemicelluloses, organic matter (OM), dry matter (DM), acid ...

  10. Induction, isolation, and characterization of aspergillus niger mutant strains producing elevated levels of beta-galactosidase.

    OpenAIRE

    Nevalainen, K M

    1981-01-01

    An Aspergillus niger mutant strain, VTT-D-80144, with an improvement of three- to fourfold in the production of extracellular beta-galactosidase was isolated after mutagenesis. The production of beta-galactosidase by this mutant was unaffected by fermentor size, and the enzyme was also suitable for immobilization.

  11. Induction, isolation, and characterization of aspergillus niger mutant strains producing elevated levels of beta-galactosidase.

    Science.gov (United States)

    Nevalainen, K M

    1981-01-01

    An Aspergillus niger mutant strain, VTT-D-80144, with an improvement of three- to fourfold in the production of extracellular beta-galactosidase was isolated after mutagenesis. The production of beta-galactosidase by this mutant was unaffected by fermentor size, and the enzyme was also suitable for immobilization. PMID:6784672

  12. In vitro evaluation of leaf extracts on the growth of Aspergillus niger infecting maize grains

    Directory of Open Access Journals (Sweden)

    Victor Okereke

    2017-04-01

    Full Text Available Mycotoxin producing fungi are important pathogens affecting stored maize grains. Conventional management strategies using synthetic chemicals are expensive, hazardous and environmentally unfriendly. This has necessitated the search for alternatives in botanicals. Market sampling of maize grains were carried out in five markets in Port Hartcourt, Nigeria, namely; Choba-main and Choba junction, Rumuosi, Aluu and Ozuoba, to evaluate the occurrence of mycotoxin-producing fungi. Studies were then carried out in the laboratory to evaluate the efficacy of water leaf extracts of Azadirachta indica, Garcina kola, Moringa oliefera, Ocimium gratissimum, Gongronema latifolium and Vernonia amygdalina at three different concentrations (2.5, 5.0 and 7.5% w/v on the prevalent fungus. Result from the market sampling showed that Aspergillus niger, Aspergillus flavus and Pennicillin sp were commonly isolated from the maize grains with Aspergillus niger having the highest occurrence. Choba markets were observed to have the highest percentage occurrence of Aspergillus niger (66-100%. Laboratory studies showed that Gongronema latifolium, effectively reduced the growth of A. niger on PDA media and leaf extract concentration at 7.5% was the most effective.

  13. Characterization of a foldase, protein disulfide isomerase A, in the protein secretory pathway of Aspergillus niger

    NARCIS (Netherlands)

    Ngiam, C.; Jeenes, D.J.; Punt, P.J.; Hondel, C.A.M.J.J. van den; Archer, D.B.

    2000-01-01

    Protein disulfide isomerase (PDI) is important in assisting the folding and maturation of secretory proteins in eukaryotes. A gene, pdiA, encoding PDIA was previously isolated from Aspergillus niger, and we report its functional characterization here. Functional analysis of PDIA showed that it

  14. Expression of the Pycnoporus cinnabarinus laccase gene in Aspergillus niger and characterization of the recombinant enzyme

    NARCIS (Netherlands)

    Record, E.; Punt, P.J.; Chamkha, M.; Labat, M.; Hondel, C.A.M.J.J. van den; Asther, M.

    2002-01-01

    Pycnoporus cinnabarinus laccase lac1 gene was overexpressed in Aspergillus niger, a well-known fungal host producing a large amount of homologous or heterologous enzymes for industrial applications. The corresponding cDNA was placed under the control of the glyceraldehyde-3-phosphate dehydrogenase

  15. Overproduction of the Aspergillus niger feruloyl esterase for pulp bleaching application

    NARCIS (Netherlands)

    Record, E.; Asther, M.; Sigoillot, C.; Pagès, S.; Punt, P.J.; Delattre, M.; Haon, M.; Hondel, C.A.M.J.J. van den; Sigoillot, J.C.; Lesage-Meessen, L.; Asther, M.

    2003-01-01

    A well-known industrial fungus for enzyme production, Aspergillus niger, was selected to produce the feruloyl esterase FAEA by homologous overexpression for pulp bleaching application. The gpd gene promoter was used to drive FAEA expression. Changing the nature and concentration of the carbon source

  16. Glucoamylase : green fluorescent protein fusions to monitor protein secretion in Aspergillus niger

    NARCIS (Netherlands)

    Gordon, C.L.; Khalaj, V.; Ram, A.F.J.; Archer, D.B.; Brookman, J.L.; Trinci, A.P.J.; Jeenes, D.J.; Doonan, J.H.; Wells, B.; Punt, P.J.; Hondel, C.A.M.J.J. van den; Robson, G.D.

    2000-01-01

    A glucoamylase: :green fluorescent protein fusion (GLA: :sGFP) was constructed which allows the green fluorescent protein to be used as an in vivo reporter of protein secretion in Aspergillus niger. Two secretory fusions were designed for secretion of GLA: :sGFP which employed slightly different

  17. Characterisation of Aspergillus niger phosphoglucose isomerase. Use for quantitative determination of erythrose 4-phosphate

    NARCIS (Netherlands)

    Ruijter, G.J.G.; Visser, J.

    1999-01-01

    Phosphoglucose isomerase (PGI) was purified from Aspergillus niger and the in vitro kinetic properties of the enzyme were related to its functioning in vivo. A new assay method was developed to study the forward reaction making use of mannitol 1-P dehydrogenase as the coupling enzyme. In this simple

  18. [Construction and application of black-box model for glucoamylase production by Aspergillus niger].

    Science.gov (United States)

    Li, Lianwei; Lu, Hongzhong; Xia, Jianye; Chu, Ju; Zhuang, Yingping; Zhang, Siliang

    2015-07-01

    Carbon-limited continuous culture was used to study the relationship between the growth of Aspergillus niger and the production of glucoamylase. The result showed that when the specific growth rate was lower than 0.068 h(-1), the production of glucoamylase was growth-associated, when the specific growth rate was higher than 0.068 h(-1), the production of glucoamylase was not growth-associated. Based on the result of continuous culture, the Monod dynamics model of glucose consumption of A. niger was constructed, Combining Herbert-Pirt equation of glucose and oxygen consumption with Luedeking-Piret equation of enzyme production, the black-box model of Aspergillus niger for enzyme production was established. The exponential fed-batch culture was designed to control the specific growth rate at 0.05 h(-1) by using this model and the highest yield for glucoamylase production by A. niger reached 0.127 g glucoamylase/g glucose. The black-box model constructed in this study successfully described the glucoamylase production by A. niger and the result of the model fitted the measured value well. The black-box model could guide the design and optimization of glucoamylase production by A. niger.

  19. Cloning and Genomic Organization of a Rhamnogalacturonase Gene from Locally Isolated Strain of Aspergillus niger.

    Science.gov (United States)

    Damak, Naourez; Abdeljalil, Salma; Taeib, Noomen Hadj; Gargouri, Ali

    2015-08-01

    The rhg gene encoding a rhamnogalacturonase was isolated from the novel strain A1 of Aspergillus niger. It consists of an ORF of 1.505 kb encoding a putative protein of 446 amino acids with a predicted molecular mass of 47 kDa, belonging to the family 28 of glycosyl hydrolases. The nature and position of amino acids comprising the active site as well as the three-dimensional structure were well conserved between the A. niger CTM10548 and fungal rhamnogalacturonases. The coding region of the rhg gene is interrupted by three short introns of 56 (introns 1 and 3) and 52 (intron 2) bp in length. The comparison of the peptide sequence with A. niger rhg sequences revealed that the A1 rhg should be an endo-rhamnogalacturonases, more homologous to rhg A than rhg B A. niger known enzymes. The comparison of rhg nucleotide sequence from A. niger A1 with rhg A from A. niger shows several base changes. Most of these changes (59 %) are located at the third base of codons suggesting maintaining the same enzyme function. We used the rhamnogalacturonase A from Aspergillus aculeatus as a template to build a structural model of rhg A1 that adopted a right-handed parallel β-helix.

  20. Overexpression of Aspergillus tubingensis faeA in protease-deficient Aspergillus niger enables ferulic acid production from plant material.

    Science.gov (United States)

    Zwane, Eunice N; Rose, Shaunita H; van Zyl, Willem H; Rumbold, Karl; Viljoen-Bloom, Marinda

    2014-06-01

    The production of ferulic acid esterase involved in the release of ferulic acid side groups from xylan was investigated in strains of Aspergillus tubingensis, Aspergillus carneus, Aspergillus niger and Rhizopus oryzae. The highest activity on triticale bran as sole carbon source was observed with the A. tubingensis T8.4 strain, which produced a type A ferulic acid esterase active against methyl p-coumarate, methyl ferulate and methyl sinapate. The activity of the A. tubingensis ferulic acid esterase (AtFAEA) was inhibited twofold by glucose and induced twofold in the presence of maize bran. An initial accumulation of endoglucanase was followed by the production of endoxylanase, suggesting a combined action with ferulic acid esterase on maize bran. A genomic copy of the A. tubingensis faeA gene was cloned and expressed in A. niger D15#26 under the control of the A. niger gpd promoter. The recombinant strain has reduced protease activity and does not acidify the media, therefore promoting high-level expression of recombinant enzymes. It produced 13.5 U/ml FAEA after 5 days on autoclaved maize bran as sole carbon source, which was threefold higher than for the A. tubingensis donor strain. The recombinant AtFAEA was able to extract 50 % of the available ferulic acid from non-pretreated maize bran, making this enzyme suitable for the biological production of ferulic acid from lignocellulosic plant material.

  1. Phytase production by Aspergillus niger CFR 335 and Aspergillus ficuum SGA 01 through submerged and solid-state fermentation.

    Science.gov (United States)

    Shivanna, Gunashree B; Venkateswaran, Govindarajulu

    2014-01-01

    Fermentation is one of the industrially important processes for the development of microbial metabolites that has immense applications in various fields. This has prompted to employ fermentation as a major technique in the production of phytase from microbial source. In this study, a comparison was made between submerged (SmF) and solid-state fermentations (SSF) for the production of phytase from Aspergillus niger CFR 335 and Aspergillus ficuum SGA 01. It was found that both the fungi were capable of producing maximum phytase on 5th day of incubation in both submerged and solid-state fermentation media. Aspergillus niger CFR 335 and A. ficuum produced a maximum of 60.6 U/gds and 38 U/gds of the enzyme, respectively, in wheat bran solid substrate medium. Enhancement in the enzyme level (76 and 50.7 U/gds) was found when grown in a combined solid substrate medium comprising wheat bran, rice bran, and groundnut cake in the ratio of 2 : 1 : 1. A maximum of 9.6 and 8.2 U/mL of enzyme activity was observed in SmF by A. niger CFR 335 and A.ficuum, respectively, when grown in potato dextrose broth.

  2. Phytase Production by Aspergillus niger CFR 335 and Aspergillus ficuum SGA 01 through Submerged and Solid-State Fermentation

    Directory of Open Access Journals (Sweden)

    Gunashree B. Shivanna

    2014-01-01

    Full Text Available Fermentation is one of the industrially important processes for the development of microbial metabolites that has immense applications in various fields. This has prompted to employ fermentation as a major technique in the production of phytase from microbial source. In this study, a comparison was made between submerged (SmF and solid-state fermentations (SSF for the production of phytase from Aspergillus niger CFR 335 and Aspergillus ficuum SGA 01. It was found that both the fungi were capable of producing maximum phytase on 5th day of incubation in both submerged and solid-state fermentation media. Aspergillus niger CFR 335 and A. ficuum produced a maximum of 60.6 U/gds and 38 U/gds of the enzyme, respectively, in wheat bran solid substrate medium. Enhancement in the enzyme level (76 and 50.7 U/gds was found when grown in a combined solid substrate medium comprising wheat bran, rice bran, and groundnut cake in the ratio of 2 : 1 : 1. A maximum of 9.6 and 8.2 U/mL of enzyme activity was observed in SmF by A. niger CFR 335 and A.ficuum, respectively, when grown in potato dextrose broth.

  3. Phytase Production by Aspergillus niger CFR 335 and Aspergillus ficuum SGA 01 through Submerged and Solid-State Fermentation

    Science.gov (United States)

    Shivanna, Gunashree B.; Venkateswaran, Govindarajulu

    2014-01-01

    Fermentation is one of the industrially important processes for the development of microbial metabolites that has immense applications in various fields. This has prompted to employ fermentation as a major technique in the production of phytase from microbial source. In this study, a comparison was made between submerged (SmF) and solid-state fermentations (SSF) for the production of phytase from Aspergillus niger CFR 335 and Aspergillus ficuum SGA 01. It was found that both the fungi were capable of producing maximum phytase on 5th day of incubation in both submerged and solid-state fermentation media. Aspergillus niger CFR 335 and A. ficuum produced a maximum of 60.6 U/gds and 38 U/gds of the enzyme, respectively, in wheat bran solid substrate medium. Enhancement in the enzyme level (76 and 50.7 U/gds) was found when grown in a combined solid substrate medium comprising wheat bran, rice bran, and groundnut cake in the ratio of 2 : 1 : 1. A maximum of 9.6 and 8.2 U/mL of enzyme activity was observed in SmF by A. niger CFR 335 and A.ficuum, respectively, when grown in potato dextrose broth. PMID:24688383

  4. Toolkit for visualization of the cellular structure and organelles in Aspergillus niger.

    Science.gov (United States)

    Buren, Emiel B J Ten; Karrenbelt, Michiel A P; Lingemann, Marit; Chordia, Shreyans; Deng, Ying; Hu, JingJing; Verest, Johanna M; Wu, Vincen; Gonzalez, Teresita J Bello; Heck, Ruben G A van; Odoni, Dorett I; Schonewille, Tom; Straat, Laura van der; Graaff, Leo H de; Passel, Mark W J van

    2014-12-19

    Aspergillus niger is a filamentous fungus that is extensively used in industrial fermentations for protein expression and the production of organic acids. Inherent biosynthetic capabilities, such as the capacity to secrete these biomolecules in high amounts, make A. niger an attractive production host. Although A. niger is renowned for this ability, the knowledge of the molecular components that underlie its production capacity, intercellular trafficking processes and secretion mechanisms is far from complete. Here, we introduce a standardized set of tools, consisting of an N-terminal GFP-actin fusion and codon optimized eforRed chromoprotein. Expression of the GFP-actin construct facilitates visualization of the actin filaments of the cytoskeleton, whereas expression of the chromoprotein construct results in a clearly distinguishable red phenotype. These experimentally validated constructs constitute the first set of standardized A. niger biomarkers, which can be used to study morphology, intercellular trafficking, and secretion phenomena.

  5. COFFEE BEAN MYCO-CONTAMINANTS AND OXALIC ACID PRODUCING ASPERGILLUS NIGER

    Directory of Open Access Journals (Sweden)

    Mohamed A. Yassin

    2015-03-01

    Full Text Available Coffee bean-contaminating fungi were determined in random samples collected in Riyadh, Kingdom of Saudi Arabia, using the direct plating technique. Forty-five samples were examined and 12 fungal species belonging to 5 genera were isolated. Aspergillus niger was the most widely distributed and most frequently isolated fungus (86.67%. The ability of the predominant fungus, A. niger, to produce oxalic acid was evaluated using high-performance liquid chromatography. About 50% of the tested A. niger isolates produced oxalic acid; the amount produced was in the range of 90–550 ppm of oxalic acid. Because A. niger was the predominant and most widely distributed toxigenic fungus in the examined samples, more efforts should be directed to minimize the risk of oxalic acid contamination of commoditized coffee beans in the Kingdom of Saudi Arabia.

  6. Advances in citric acid fermentation by Aspergillus niger: biochemical aspects, membrane transport and modeling.

    Science.gov (United States)

    Papagianni, Maria

    2007-01-01

    Citric acid is regarded as a metabolite of energy metabolism, of which the concentration will rise to appreciable amounts only under conditions of substantive metabolic imbalances. Citric acid fermentation conditions were established during the 1930s and 1940s, when the effects of various medium components were evaluated. The biochemical mechanism by which Aspergillus niger accumulates citric acid has continued to attract interest even though its commercial production by fermentation has been established for decades. Although extensive basic biochemical research has been carried out with A. niger, the understanding of the events relevant for citric acid accumulation is not completely understood. This review is focused on citric acid fermentation by A. niger. Emphasis is given to aspects of fermentation biochemistry, membrane transport in A. niger and modeling of the production process.

  7. VeA of Aspergillus niger increases spore dispersing capacity by impacting conidiophore architecture.

    Science.gov (United States)

    Wang, Fengfeng; Dijksterhuis, Jan; Wyatt, Timon; Wösten, Han A B; Bleichrodt, Robert-Jan

    2015-01-01

    Aspergillus species are highly abundant fungi worldwide. Their conidia are among the most dominant fungal spores in the air. Conidia are formed in chains on the vesicle of the asexual reproductive structure called the conidiophore. Here, it is shown that the velvet protein VeA of Aspergillus niger maximizes the diameter of the vesicle and the spore chain length. The length and width of the conidiophore stalk and vesicle were reduced nearly twofold in a ΔveA strain. The latter implies a fourfold reduced surface area to develop chains of spores. Over and above this, the conidial chain length was approximately fivefold reduced. The calculated 20-fold reduction in formation of conidia by ΔveA fits the 8- to 17-fold decrease in counted spore numbers. Notably, morphology of the ΔveA conidiophores of A. niger was very similar to that of wild-type Aspergillus sydowii. This suggests that VeA is key in conidiophore architecture diversity in the fungal kingdom. The finding that biomass formation of the A. niger ΔveA strain was reduced twofold shows that VeA not only impacts dispersion capacity but also colonization capacity of A. niger.

  8. The Aspergillus niger growth on the treated concrete substrate using variable antifungals

    Science.gov (United States)

    Parjo, U. K.; Sunar, N. M.; Leman, A. M.; Gani, P.; Embong, Z.; Tajudin, S. A. A.

    2016-11-01

    The aim of this study was to evaluate the Aspergillus niger (A. niger) growth on substrates after incorporates with different compounds of antifungals which is normally used in food industry. The antifungals named as potassium sorbate (PS), calcium benzoate (CB) and zinc salicylate (ZS) were applied on concrete substrate covered with different wall finishing such as acrylic paint (AP), glycerol based paint (GBP), thin wallpaper (THIN) and thick wallpaper (THICK). The concrete substrate were inoculated with spore suspension, incubated at selected temperature (30oC) and relative humidity (90%)in plant growth chamber. The observations were done from the Day 3 until Day 27. The results showed that the growth of the A. niger for concrete treated by PS for AP, GBP, THIN, and THICK were 64%, 32%, 11% and 100%, respectively. Meanwhile for CB, the growth of A. niger on AP, GBP, THIN, and THICK were 100%, 12%, 41%, and 13%, respectively. Similarly, treated concrete by ZS revealed that the growth of A. niger on the same substrate cover were 33%, 47%, 40%, and 39%, respectively. The results obtained in this study provide a valuable knowledge on the abilities of antifungals to remediate A. niger that inoculated on the concrete substrate. Consequently, this study proved that the PS covering with THIN more efficiency compares CB and ZS to prevent A. niger growth.

  9. Citric acid production by Aspergillus niger on wet corn distillers grains.

    Science.gov (United States)

    Xie, G; West, T P

    2006-09-01

    To determine which citric acid-producing strain of Aspergillus niger utilized wet corn distillers grains most effectively to produce citric acid. Citric acid and biomass production by the fungal strains were analysed on the untreated grains or autoclaved grains using an enzyme assay and a gravimetric method respectively. Fungal citric acid production on the grains was found to occur on the untreated or autoclaved grains. The highest citric acid level on the grains was produced by A. niger ATCC 9142. The autoclaved grains supported less citric acid production by the majority of strains screened. Biomass production by the fungal strains on the untreated or autoclaved grains was quite similar. The highest citric acid yields for A. niger ATCC 9142, ATCC 10577, ATCC 11414, ATCC 12846 and ATCC 26550 were found on the untreated grains. Treatment of the grains had little effect on citric acid yields based on reducing sugars consumed by A. niger ATCC 9029 and ATCC 201122. It is feasible for citric acid-producing strains of A. niger to excrete citric acid on wet corn distillers grains whether the grains are treated or untreated. The most effective citric acid-producing strain of A. niger was ATCC 9142. The study shows that the ethanol processing co-product wet corn distillers grains could be utilized as a substrate for the commercial production of citric acid by A. niger without treatment of the grains.

  10. Biosynthesis of silver nanoparticles by Aspergillus niger , Fusarium ...

    African Journals Online (AJOL)

    ... scanning electron microscope (SEM). Results indicate the synthesis of silver nanoparticles in the reaction mixture. The synthesis of nanoparticles would be suitable for developing a microbial nanotechnology biosynthesis process for mass scale production. Keywords: Silver nanoparticles, biosynthesis, fungi, Aspergillus.

  11. Constitutive expression of fluorescent protein by Aspergillus var. niger and Aspergillus carbonarius to monitor fungal colonization in maize plants.

    Science.gov (United States)

    Palencia, Edwin Rene; Glenn, Anthony Elbie; Hinton, Dorothy Mae; Bacon, Charles Wilson

    2013-09-01

    Aspergillus niger and Aspergillus carbonarius are two species in the Aspergillus section Nigri (black-spored aspergilli) frequently associated with peanut (Arachis hypogea), maize (Zea mays), and other plants as pathogens. These infections are symptomless and as such are major concerns since some black aspergilli produce important mycotoxins, ochratoxins A, and the fumonisins. To facilitate the study of the black aspergilli-maize interactions with maize during the early stages of infections, we developed a method that used the enhanced yellow fluorescent protein (eYFP) and the monomeric red fluorescent protein (mRFP1) to transform A. niger and A. carbonarius, respectively. The results were constitutive expressions of the fluorescent genes that were stable in the cytoplasms of hyphae and conidia under natural environmental conditions. The hyphal in planta distribution in 21-day-old seedlings of maize were similar wild type and transformants of A. niger and A. carbonarius. The in planta studies indicated that both wild type and transformants internally colonized leaf, stem and root tissues of maize seedlings, without any visible disease symptoms. Yellow and red fluorescent strains were capable of invading epidermal cells of maize roots intercellularly within the first 3 days after inoculation, but intracellular hyphal growth was more evident after 7 days of inoculation. We also tested the capacity of fluorescent transformants to produce ochratoxin A and the results with A. carbonarius showed that this transgenic strain produced similar concentrations of this secondary metabolite. This is the first report on the in planta expression of fluorescent proteins that should be useful to study the internal plant colonization patterns of two ochratoxigenic species in the Aspergillus section Nigri. © 2013.

  12. Citric acid production by selected mutants of Aspergillus niger from cane molasses.

    Science.gov (United States)

    Ikram-Ul, Haq; Ali, Sikander; Qadeer, M A; Iqbal, Javed

    2004-06-01

    The present investigation deals with citric acid production by some selected mutant strains of Aspergillus niger from cane molasses in 250 ml Erlenmeyer flasks. For this purpose, a conidial suspension of A. niger GCB-75, which produced 31.1 g/l citric acid from 15% (w/v) molasses sugar, was subjected to UV-induced mutagenesis. Among the 3 variants, GCM-45 was found to be a better producer of citric acid (50.0 +/- 2a) and it was further improved by chemical mutagenesis using N-methyl, N-nitro-N-nitroso-guanidine (MNNG). Out of 3,2-deoxy-D-glucose resistant variants, GCMC-7 was selected as the best mutant, which produced 96.1 +/- 1.5 g/l citric acid 168 h after fermentation of potassium ferrocyanide and H2SO4 pre-treated blackstrap molasses in Vogel's medium. On the basis of kinetic parameters such as volumetric substrate uptake rate (Qs), and specific substrate uptake rate (qs), the volumetric productivity, theoretical yield and specific product formation rate, it was observed that the mutants were faster growing organisms and produced more citric acid. The mutant GCMC-7 has greater commercial potential than the parental strain with regard to citrate synthase activity. The addition of 2.0 x 10(-5) M MgSO4 x 5H2O into the fermentation medium reduced the Fe2+ ion concentration by counter-acting its deleterious effect on mycelial growth. The magnesium ions also induced a loose-pelleted form of growth (0.6 mm, diameter), reduced the biomass concentration (12.5 g/l) and increased the volumetric productivity of citric acid monohydrate (113.6 +/- 5 g/l). Copyright 2003 Elsevier Ltd.

  13. Performance of Aspergillus niger Cultivation in Geometrically Dissimilar Bioreactors Evaluated on the Basis of Morphological Analyses

    Directory of Open Access Journals (Sweden)

    M. A. Priede

    2002-01-01

    Full Text Available The growth of Aspergillus niger, citric acid production and mycelia morphology changes were compared under different mixing conditions in bioreactors with two types of stirrers: Rushton turbine stirrers (RTS1 or RTS2 and axial counterflow stirrers (ACS1 or ACS2. The characteristics of growth, productivity and morphology varied with the mixing system and the applied agitation regime. In the first series of experiments, the flow characteristics of Aspergillus niger broth under different mixing conditions were analysed in a model bioreactor using RTS1 and ACS1. The kinetic energy E of flow fluctuations was measured in gassed and ungassed water and fermentation broth systems using a stirring intensity measuring device (SIMD-f1. The difference of energy E values at different points was more pronounced in the bioreactor with RTS1 than in the case of ACS1. High viscous A. niger broths provided higher energy E values in comparison with water. It was observed that the Aspergillus niger growth rate and citric acid synthesis rate decreased at very high energy E values, the behaviour obviously being connected with the influence of the irreversible shear stress on the mycelial morphology. In the second series of experiments, a higher citric acid yield was achieved in the case of ACS2 at a power input approximately twice lower than in the case of RTS2. Morphological characterization of A. niger pellets was carried out by the image analysis method. ACS2 provided the development of morphology, where pellets and cores had larger area, perimeter and diameter, and the annular region of pellets was looser and more »hairy« in comparison with the case of RTS2. The pellets from the fermentation with RTS2 were smaller, denser, with shorter hyphae in the annular region of pellets, and the broth was characterized by a higher percentage of diffuse mycelia. Power input studies of RTS2 and ACS2 were made at different agitator rotation speeds and gas flow rates using water

  14. Recombinant hepatitis B surface antigen production in Aspergillus niger: evaluating the strategy of gene fusion to native glucoamylase

    CSIR Research Space (South Africa)

    James, ER

    2012-10-01

    Full Text Available Microbiology and Biotechnology October 2012/ Vol. 96, No.2 Recombinant hepatitis B surface antigen production in Aspergillus niger: evaluating the strategy of gene fusion to native glucoamylase ER James a,c & WH van Zyl b & PJ van Zyl c & JF Görgens..., Pretoria 0001, South Africa Abstract This study demonstrates the potential of Aspergillus niger as a candidate expression system for virus- like particle production using gene fusion. Hepatitis B surface antigen (HBsAg) production, targeted...

  15. Production of Antifungal Chitinase by Aspergillus niger LOCK 62 and Its Potential Role in the Biological Control

    OpenAIRE

    Brzezinska, Maria Swiontek; Jankiewicz, Urszula

    2012-01-01

    Aspergillus niger LOCK 62 produces an antifungal chitinase. Different sources of chitin in the medium were used to test the production of the chitinase. Chitinase production was most effective when colloidal chitin and shrimp shell were used as substrates. The optimum incubation period for chitinase production by Aspergillus niger LOCK 62 was 6 days. The chitinase was purified from the culture medium by fractionation with ammonium sulfate and affinity chromatography. The molecular mass of the...

  16. Systemic analysis of the response of Aspergillus niger to ambient pH

    DEFF Research Database (Denmark)

    Andersen, Mikael Rørdam; Lehmann, Linda Olkjær; Nielsen, Jens

    2009-01-01

    efficient acidification as well as production process applicable knowledge on the transcriptional regulation of pH response in the industrially important A. niger. It has also made clear that filamentous fungi have evolved to employ several offensive strategies for out-competing rival organisms.......Background: The filamentous fungus Aspergillus niger is an exceptionally efficient producer of organic acids, which is one of the reasons for its relevance to industrial processes and commercial importance. While it is known that the mechanisms regulating this production are tied to the levels...

  17. Microbial transformation of artemisinin to 5-hydroxyartemisinin by Eurotium amstelodami and Aspergillus niger.

    Science.gov (United States)

    Parshikov, Igor A; Miriyala, Brushapathy; Muraleedharan, Kannoth M; Avery, Mitchell A; Williamson, John S

    2006-05-01

    Transformation of the anti-malarial drug artemisinin by the fungi Eurotium amstelodami and Aspergillus niger were investigated. Cultures were grown in sucrose/malt broth with artemisinin for 14 days and extracted with ethyl acetate. Extracts were characterized by liquid chromatography. Two metabolites from each fungal extract were isolated and identified using mass spectrometry and nuclear magnetic resonance. 5Beta-hydroxyartemisinin and 7beta-hydroxyartemisinin were isolated in 63 and 32% yields, respectively, from the extract of E. amstelodami, and 80 and 19%, respectively, from the extract of A. niger.

  18. Molecular genetic analysis of vesicular transport in Aspergillus niger reveals partial conservation of the molecular mechanism of exocytosis in fungi

    NARCIS (Netherlands)

    Kwon, M.J.; Arentshorst, M.; Fiedler, M.; Groen,F.L.M. de; Punt, P.J.; Meyer, V.; Ram, A.F.J

    2014-01-01

    The filamentous fungus Aspergillus niger is an industrially exploited protein expression platform, well known for its capacity to secrete high levels of proteins. To study the process of protein secretion in A. niger, we established a GFP-v-SNARE reporter strain in which the trafficking and dynamics

  19. Lactobacillus brevis-based bioingredient inhibits Aspergillus niger growth on pan bread

    Directory of Open Access Journals (Sweden)

    Mariaelena Di Biase

    2014-11-01

    Full Text Available Bread shelf life is generally compromised by fungi mainly belonging to Aspergillus and Penicillium genera, which colonise the surface of the product within few days from the production. The aim of this study was to select a Lactobacillus brevis-based bioingredient (LbBio able to inhibit the growth of Aspergillus niger ITEM5132 on pan bread in order to prolong its shelf life. Four LbBio formulations, obtained by growing a selected L. brevis strain in a flour-based medium containing different carbon sources or acid precursors (fructose, LbBio1; fructose and maltose, LbBio2; α-chetoglutaric acid, LbBio3; short-chain fructooligosaccharides, LbBio4, were evaluated for their content of organic acids (lactic, acetic, propionic, phenyllactic, 4-hydroxy-phenyllactic, valeric, isovaleric acids. The LbBio formulations were applied in yeast-leavened bread during bread-making trials and the resulting products were inoculated after baking with A. niger spore’s suspension and the fungal growth was monitored during storage (25°C for 6 days. The formulation showing the highest inhibitory activity was separated by ultra-filtration method, and whole and fractions obtained were evaluated for their in vitro activity. The fraction showing the highest activity was further separated by gel-filtration and the resulting products were investigated for their protein content and in vitro inhibition. The results from the bread-making trials performed using different formulations of LbBio showed a delay in fungal growth (1 day respect to the bread not containing the bioingredient (control or including calcium propionate (0.3% w/w. The formulation LbBio2, prepared with fructose and maltose 1% (w/vol, contained the highest amount of total organic acids, including phenyllactic and hydroxyl-phenyllactic acids, and reduced the visual spoilage of bread. This formulation was separated by ultra-filtration and fractions containing metabolites with molecular weight higher than 30 k

  20. Pectinolytic complex production by Aspergillus niger URM 4645 ...

    African Journals Online (AJOL)

    SAM

    2014-08-06

    Aug 6, 2014 ... presenting a pectinolytic index of 0.51 (Table 2). Bezerra et al. (2012) reported that among 24 endophytic fungal isolates of Opuntia ficus-indica (forage cactus), only A. japonicus and Penicillium glandicola showed pectinolytic activity. Production of pectinolytic enzymes. A. niger URM 4645 was the isolate ...

  1. Assessment of Aspergillus niger biofilm growth kinetics in ...

    African Journals Online (AJOL)

    Jane

    2011-10-12

    Oct 12, 2011 ... other hand, A. niger biofilm growth followed a logistic model having higher maximal specific growth rate than submerged cultures (0.061 ... applications in the food industry, beverages, textiles, agriculture, pulp and paper and ..... curves showed the same trend over time (Figure 4A). However, in submerged ...

  2. Gluconate formation and polyol metabolism in Aspergillus niger

    NARCIS (Netherlands)

    Witteveen, C.F.B.

    1993-01-01

    The capacity of A.niger to accumulate metabolites is remarkable. Under all conditions polyols accumulate in the cell and when mycelium in later developmental stages is considered, depending on the carbon source, aeration and external pH, polyols

  3. Evaluation of xylanases from Aspergillus niger and Trichoderma sp ...

    African Journals Online (AJOL)

    Jane

    2011-08-17

    Aug 17, 2011 ... properties of the gluten make the dough more viscous than elastic, which does not provide increased volume or ... the flour and the effect of enzymes in it. The method recommended by AACC 54-10 (1995) ... Table 1, the xylanase from A. niger did not affect the stability of the flour. Xylanase derived from the.

  4. Effect of temperature, water activity, and pH on growth and production of ochratoxin A by Aspergillus niger and Aspergillus carbonarius from Brazilian grapes.

    Science.gov (United States)

    Passamani, Fabiana Reinis Franca; Hernandes, Thais; Lopes, Noelly Alves; Bastos, Sabrina Carvalho; Santiago, Wilder Douglas; Cardoso, Maria das Graças; Batista, Luís Roberto

    2014-11-01

    The growth of ochratoxigenic fungus and the presence of ochratoxin A (OTA) in grapes and their derivatives can be caused by a wide range of physical, chemical, and biological factors. The determination of interactions between these factors and fungal species from different climatic regions is important in designing models for minimizing the risk of OTA in wine and grape juice. This study evaluated the influence of temperature, water activity (aw), and pH on the development and production of OTA in a semisynthetic grape culture medium by Aspergillus carbonarius and Aspergillus niger strains. To analyze the growth conditions and production of OTA, an experimental design was conducted using response surface methodology as a tool to assess the effects of these abiotic variables on fungal behavior. A. carbonarius showed the highest growth at temperatures from 20 to 33°C, aw between 0.95 and 0.98, and pH levels between 5 and 6.5. Similarly, for A. niger, temperatures between 24 and 37°C, aw greater than 0.95, and pH levels between 4 and 6.5 were optimal. The greatest toxin concentrations for A. carbonarius and A. niger (10 μg/g and 7.0 μg/g, respectively) were found at 15°C, aw 0.99, and pH 5.35. The lowest pH was found to contribute to greater OTA production. These results show that the evaluated fungi are able to grow and produce OTA in a wide range of temperature, aw, and pH. However, the optimal conditions for toxin production are generally different from those optimal for fungal growth. The knowledge of optimal conditions for fungal growth and production of OTA, and of the stages of cultivation in which these conditions are optimal, allows a more precise assessment of the potential risk to health from consumption of products derived from grapes.

  5. Korean Ginseng Berry Fermented by Mycotoxin Non-producing Aspergillus niger and Aspergillus oryzae: Ginsenoside Analyses and Anti-proliferative Activities.

    Science.gov (United States)

    Li, Zhipeng; Ahn, Hyung Jin; Kim, Nam Yeon; Lee, Yu Na; Ji, Geun Eog

    2016-01-01

    To transform ginsenosides, Korean ginseng berry (KGB) was fermented by mycotoxin non-producing Aspergillus niger and Aspergillus oryzae. Changes of ginsenoside profile and anti-proliferative activities were observed. Results showed that A. niger tended to efficiently transform protopanaxadiol (PPD) type ginsenosides such as Rb1, Rb2, Rd to compound K while A. oryzae tended to efficiently transform protopanaxatriol (PPT) type ginsenoside Re to Rh1 via Rg1. Butanol extracts of fermented KGB showed high cytotoxicity on human adenocarcinoma HT-29 cell line and hepatocellular carcinoma HepG2 cell line while that of unfermented KGB showed little. The minimum effective concentration of niger-fermented KGB was less than 2.5 µg/mL while that of oryzae-fermented KGB was about 5 µg/mL. As A. niger is more inclined to transform PPD type ginsenosides, niger-fermented KGB showed stronger anti-proliferative activity than oryzae-fermented KGB.

  6. Expression of Lactate Dehydrogenase in Aspergillus niger for L-Lactic Acid Production

    Science.gov (United States)

    Dave, Khyati K.; Punekar, Narayan S.

    2015-01-01

    Different engineered organisms have been used to produce L-lactate. Poor yields of lactate at low pH and expensive downstream processing remain as bottlenecks. Aspergillus niger is a prolific citrate producer and a remarkably acid tolerant fungus. Neither a functional lactate dehydrogenase (LDH) from nor lactate production by A. niger is reported. Its genome was also investigated for the presence of a functional ldh. The endogenous A. niger citrate synthase promoter relevant to A. niger acidogenic metabolism was employed to drive constitutive expression of mouse lactate dehydrogenase (mldhA). An appraisal of different branches of the A. niger pyruvate node guided the choice of mldhA for heterologous expression. A high copy number transformant C12 strain, displaying highest LDH specific activity, was analyzed under different growth conditions. The C12 strain produced 7.7 g/l of extracellular L-lactate from 60 g/l of glucose, in non-neutralizing minimal media. Significantly, lactate and citrate accumulated under two different growth conditions. Already an established acidogenic platform, A. niger now promises to be a valuable host for lactate production. PMID:26683313

  7. Production of Fumonisin B-2 and B-4 by Aspergillus niger on Grapes and Raisins

    DEFF Research Database (Denmark)

    Mogensen, Jesper Mølgaard; Frisvad, Jens Christian; Thrane, Ulf

    2010-01-01

    The recent discovery of fumonisin production in Aspergillus niger, raises concerns about the presence of these mycotoxins in grapes and raisins as well as other commodities where A. niger is a frequent contaminant. Here we investigate the potential production of fumonisins in A. niger cultured...... on grapes and raisins. Sixty-six A. niger, 4 A. tubingensis, and 16 A. acidus strains isolated from raisins were tested for fumonisin production on laboratory media. Neither A. tubingensis nor A. acidus strains produced fumonisins, but 77% of A. niger strains did. None of the strains produced ochratoxin A....... Ten selected fumonisin producing A. niger strains were further able to produce fumonisin B2 and fumonisin B4 on grapes in the range 171−7841 μg fumonisin B2/kg and 14−1157 μg fumonisin B4/kg. Four selected strains were able to produce fumonisin B2 (5−6476 μg/kg) and fumonisin B4 (12−672 μg...

  8. Expression of Lactate Dehydrogenase in Aspergillus niger for L-Lactic Acid Production.

    Science.gov (United States)

    Dave, Khyati K; Punekar, Narayan S

    2015-01-01

    Different engineered organisms have been used to produce L-lactate. Poor yields of lactate at low pH and expensive downstream processing remain as bottlenecks. Aspergillus niger is a prolific citrate producer and a remarkably acid tolerant fungus. Neither a functional lactate dehydrogenase (LDH) from nor lactate production by A. niger is reported. Its genome was also investigated for the presence of a functional ldh. The endogenous A. niger citrate synthase promoter relevant to A. niger acidogenic metabolism was employed to drive constitutive expression of mouse lactate dehydrogenase (mldhA). An appraisal of different branches of the A. niger pyruvate node guided the choice of mldhA for heterologous expression. A high copy number transformant C12 strain, displaying highest LDH specific activity, was analyzed under different growth conditions. The C12 strain produced 7.7 g/l of extracellular L-lactate from 60 g/l of glucose, in non-neutralizing minimal media. Significantly, lactate and citrate accumulated under two different growth conditions. Already an established acidogenic platform, A. niger now promises to be a valuable host for lactate production.

  9. Effect of different polyphenol sources on the efficiency of ellagic acid release by Aspergillus niger.

    Science.gov (United States)

    Sepúlveda, Leonardo; de la Cruz, Reynaldo; Buenrostro, José Juan; Ascacio-Valdés, Juan Alberto; Aguilera-Carbó, Antonio Francisco; Prado, Arely; Rodríguez-Herrera, Raúl; Aguilar, Cristóbal Noé

    2016-01-01

    Fungal hydrolysis of ellagitannins produces hexahydroxydiphenic acid, which is considered an intermediate molecule in ellagic acid release. Ellagic acid has important and desirable beneficial health properties. The aim of this work was to identify the effect of different sources of ellagitannins on the efficiency of ellagic acid release by Aspergillus niger. Three strains of A. niger (GH1, PSH and HT4) were assessed for ellagic acid release from different polyphenol sources: cranberry, creosote bush, and pomegranate used as substrate. Polyurethane foam was used as support for solid-state culture in column reactors. Ellagitannase activity was measured for each of the treatments. Ellagic acid was quantified by high performance liquid chromatography. When pomegranate polyphenols were used, a maximum value of ellagic acid (350.21 mg/g) was reached with A. niger HT4 in solid-state culture. The highest amount of ellagitannase (5176.81 U/l) was obtained at 8h of culture when cranberry polyphenols and strain A. niger PSH were used. Results demonstrated the effect of different polyphenol sources and A. niger strains on ellagic acid release. It was observed that the best source for releasing ellagic acid was pomegranate polyphenols and A. niger HT4 strain, which has the ability to degrade these compounds for obtaining a potent bioactive molecule such as ellagic acid. Copyright © 2015 Asociación Argentina de Microbiología. Publicado por Elsevier España, S.L.U. All rights reserved.

  10. Dephosphorylation of phytate by using the Aspergillus niger phytase with a high affinity for phytate.

    Science.gov (United States)

    Nagashima, T; Tange, T; Anazawa, H

    1999-10-01

    A phytase (EC 3.1.3.8) with a high affinity for phytic acid was found in Aspergillus niger SK-57 and purified to homogeneity in four steps by using ion-exchange chromatography (two types), gel filtration, and chromatofocusing. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of the purified enzyme gave a single stained band at a molecular mass of approximately 60 kDa. The Michaelis constant of the enzyme for phytic acid (18.7 +/- 4.6 microM) was statistically analyzed. In regard to the orthophosphate released from phytic acid, a significant difference between a low K(m) phytase from A. niger SK-57 and a high K(m) phytase from Aspergillus ficuum was recognized.

  11. The promoter of the glucoamylase-encoding gene of Aspergillus niger functions in Ustilago maydis

    Energy Technology Data Exchange (ETDEWEB)

    Smith, T.L. (Dept. of Agriculture, Madison, WI (United States) Univ. of Wisconsin, Madison (United States)); Gaskell, J.; Cullen, D. (Dept. of Agriculture, Madison, WI (United States)); Berka, R.M.; Yang, M.; Henner, D.J. (Genentech Inc., San Francisco, CA (United States))

    1990-01-01

    Promoter sequences from the Aspergillus niger glucoamylase-encoding gene (glaA) were linked to the bacterial hygromycin (Hy) phosphotransferase-encoding gene (hph) and this chimeric marker was used to select Hy-resistant (Hy[sup R]) Ustilago maydis transformants. This is an example of an Ascomycete promoter functioning in a Basidiomycete. Hy[sup R] transformants varied with respect to copy number of integrated vector, mitotic stability, and tolerance to Hy. Only 216 bp of glaA promoter sequence is required for expression in U. maydis but this promoter is not induced by starch as it is in Aspergillus spp. The transcription start points are the same in U. maydis and A. niger.

  12. Prospecting for the incidence of genes involved in ochratoxin and fumonisin biosynthesis in Brazilian strains of Aspergillus niger and Aspergillus welwitschiae.

    Science.gov (United States)

    Massi, Fernanda Pelisson; Sartori, Daniele; de Souza Ferranti, Larissa; Iamanaka, Beatriz Thie; Taniwaki, Marta Hiromi; Vieira, Maria Lucia Carneiro; Fungaro, Maria Helena Pelegrinelli

    2016-03-16

    Aspergillus niger "aggregate" is an informal taxonomic rank that represents a group of species from the section Nigri. Among A. niger "aggregate" species Aspergillus niger sensu stricto and its cryptic species Aspergillus welwitschiae (=Aspergillus awamori sensu Perrone) are proven as ochratoxin A and fumonisin B2 producing species. A. niger has been frequently found in tropical and subtropical foods. A. welwitschiae is a new species, which was recently dismembered from the A. niger taxon. These species are morphologically very similar and molecular data are indispensable for their identification. A total of 175 Brazilian isolates previously identified as A. niger collected from dried fruits, Brazil nuts, coffee beans, grapes, cocoa and onions were investigated in this study. Based on partial calmodulin gene sequences about one-half of our isolates were identified as A. welwitschiae. This new species was the predominant species in onions analyzed in Brazil. A. niger and A. welwitschiae differ in their ability to produce ochratoxin A and fumonisin B2. Among A. niger isolates, approximately 32% were OTA producers, but in contrast only 1% of the A. welwitschiae isolates revealed the ability to produce ochratoxin A. Regarding fumonisin B2 production, there was a higher frequency of FB2 producing isolates in A. niger (74%) compared to A. welwitschiae (34%). Because not all A. niger and A. welwitschiae strains produce ochratoxin A and fumonisin B2, in this study a multiplex PCR was developed for detecting the presence of essential genes involved in ochratoxin (polyketide synthase and radHflavin-dependent halogenase) and fumonisin (α-oxoamine synthase) biosynthesis in the genome of A. niger and A. welwitschiae isolates. The frequency of strains harboring the mycotoxin genes was markedly different between A. niger and A. welwitschiae. All OTA producing isolates of A. niger and A. welwitschiae showed in their genome the pks and radH genes, and 95.2% of the nonproducing

  13. Simultaneous amyloglucosidase and exo-polygalacturonase production by Aspergillus niger using solid-state fermentation

    Directory of Open Access Journals (Sweden)

    Jorge Alberto Vieira Costa

    2007-09-01

    Full Text Available Amyloglucosidase (AMG and exo-polygalacturonase (exo-PG were simultaneously produced by two different strains of Aspergillus niger in solid-state fermentation (SSF using defatted rice-bran as substrate. The effect of Aspergillus niger strain (t0005/007-2 and/or CCT 3312, inoculum type (spore suspension or fermented bran and addition of inducers (pectin and/or starch to the culture media was studied using a 3² x 2¹ factorial experimental design. The production of AMG and exo-PG was significantly affected by fungal strain and inoculum type but inducers had no effect. The maximum yields obtained were 1310 U/g dm for AMG using a spore suspension of A. niger CCT 3312 and 50.2 U/g dm for exo-PG production, using A. niger t0005/007-2 and fermented bran as inoculum. The yields obtained represented acceptable values in comparison with data available in the literature and indicated that defatted rice-bran was a good nutrient source.As enzimas amiloglicosidase (AMG e exo-poligalacturonase (exo-PG foram produzidas simultaneamente por duas cepas de Aspergillus niger, através de fermentação em estado sólido usando farelo de arroz desengordurado como substrato. Foram avaliados os efeitos da cepa de Aspergillus niger, tipo de inóculo e adição de indutores no meio de cultura, utilizando-se um planejamento experimental fracionário 3² x 2¹. O máximo rendimento obtido foi 1310 U/g ms para a produção de AMG e 50,2 U/g ms para a exo-PG. Comparando-se estes resultados com dados da literatura pode-se dizer que os rendimentos obtidos foram aceitáveis e indicam que o farelo de arroz desengordurado é uma boa fonte de nutrientes. A produção de AMG e exo-PG foi significativamente afetada pelas variáveis cepa de A. niger e tipo de inóculo, enquanto a variável indutor não apresentou influência significativa na produção destas enzimas.

  14. Lipase production by recombinant strains of Aspergillus niger expressing a lipase-encoding gene from Thermomyces lanuginosus

    DEFF Research Database (Denmark)

    Prathumpai, Wai; Flitter, S.J.; Mcintyre, Mhairi

    2004-01-01

    Two recombinant strains of Aspergillus niger (NW 297-14 and NW297-24) producing a heterologous lipase from Thermomyces lanuginosus were constructed. The heterologous lipase was expressed using the TAKA amylase promoter from Aspergillus oryzae. The production kinetics of the two strains on different...

  15. The kinetics of glucose production from rice straw by Aspergillus niger

    African Journals Online (AJOL)

    In this investigation, glucose was produced from rice straw using cells of Aspergillus niger, isolated from maize grain. Glucose yield was found to increase from 43 to 87% as the rice straw particle size decreased from 425 to 75 ìm, while the optimal temperature and pH were found within the range of 45 - 50°C and 4.5 - 5 ...

  16. OPTIMIZATION OF EXTRACELLULAR TANNASE PRODUCTION BY ASPERGILLUS NIGER VAN TIEGHEM USING RESPONSE SURFACE METHODOLOGY

    OpenAIRE

    Hamada Abou-Bakr; Malak El-Sahn; Amr El-Banna

    2013-01-01

    Response surface methodology (RSM) was used to optimize the production of tannase by a newly isolate of Aspergillus niger Van Tieghem using rotatable central composite design (RCCD). This statistical optimization process was carried out involving four of quantitative growth parameters (variables), namely tannic acid concentration, nitrogen source concentration, initial pH of the medium and inoculum size. A mathematical model expressing the production process of tannase by submerged fermentati...

  17. Bioprocessing of citrus waste peel for induced pectinase production by Aspergillus niger; its purification and characterization

    OpenAIRE

    Ahmed, Ishtiaq; Zia, Muhammad Anjum; Hussain, Muhammad Azhar; Akram, Zain; Naveed, Muhammad Tahir; Nowrouzi, Azin

    2016-01-01

    Agro-industrial residues are primarily composed of complex polysaccharides that strengthen microbial growth for the production of industrially important enzymes. Pectinases are one of the most widely disseminated enzymes in bacteria, fungi and plants. Czapeck media supplemented with orange waste peel as carbon source under submerged fermentation process Aspergillus niger presenting the preeminent enzymatic production. On partial optimization culture showed the maximum enzyme yield (117.1 ± 3....

  18. Optimization of extraction of β-endoglucanase from the fermented bran of Aspergillus niger

    OpenAIRE

    Chandra, M. Subhosh; Viswanath, Buddolla; Reddy, B. Rajasekhar

    2010-01-01

    A local isolate of Aspergillus niger was cultivated under optimal growth conditions on wheat bran in solid state fermentation. β-endoglucanase from fermented bran was separately extracted with different solvents to test recovery of enzyme. Among solvents tested, distilled water served the best leachate. Conditions were further optimized with this leachate. Two washes of fermented bran with the leachate for 30 min each under shaking conditions in a ratio of 1 g of wheat bran: 4 ml of distilled...

  19. Optimization and stability of glucoamylase production by recombinant strains of Aspergillus niger in chemostat culture

    NARCIS (Netherlands)

    Withers, J.M.; Swift, R.J.; Wiebe, M.G.; Robson, G.D.; Punt, P.J.; Hondel, C.A.M.J.J. van den

    1998-01-01

    When grown on a medium containing 5 g maltodextrin L-1, Aspergillus niger transformant N402[pAB6-10]B1, which has an additional 20 copies of the glucoamylase (glaA) gene, produced 320 ± 8 mg (mean ± S.E.) glucoamylase (GAM) L-1 in batch culture and 373 ± 9 mg GAM L-1 in maltodextrin- limited

  20. Selection of 5-fluorocytosine-resistant mutants from an Aspergillus niger citric acid-producing strain

    Directory of Open Access Journals (Sweden)

    Conte Ana Paula de F.

    2003-01-01

    Full Text Available Mutants of Aspergillus niger N402, induced by UV mutagenesis, were selected and tested for resistance or sensitivity to 5-fluorocytosine. Some mutants showed increased citric acid production, which did not correlate with the intracellular amount of protein or ammonium ion. The resistance to 5-fluorocytosine proved to be a rational approach for isolation of new mutants with improved production of citric acid. The best mutant (FR13 accumulated double the amount of citric acid produced by the parental strain.

  1. Biotransformations of Imbricatolic Acid by Aspergillus niger and Rhizopus nigricans Cultures

    OpenAIRE

    Cristina Theoduloz; Jaime A Rodríguez; Marcela Kurina; Carlos Aranda; Guillermo Schmeda-Hirschmann

    2007-01-01

    Microbial transformation of imbricatolic acid (1) by Aspergillus niger afforded 1α-hydroxyimbricatolic acid (2), while transformation with Rhizopus nigricans yielded 15-hydroxy-8,17-epoxylabdan-19-oic acid (3). When the diterpene 1 was added to a Cunninghamella echinulata culture, the main products were the microbial metabolites mycophenolic acid (4) and its 3-hydroxy derivative 5. All the structures were elucidated by spectroscopic methods. The cytotoxicity of these compounds towards huma...

  2. Aspergillus niger β-galactosidase production by yeast in a continuous high cell density reactor

    OpenAIRE

    Domingues, Lucília; Lima, Nelson; Teixeira, J. A.

    2005-01-01

    The continuous production of extracellular heterologous beta-galactosidase by a recombinant flocculating Saccharomyces cerevisiae, expressing the lacA gene (coding for beta-galactosidase) of Aspergillus niger was investigated. A continuous operation was run in a 6.51 airlift bioreactor with a concentric draft tube using lactose as substrate. Data on the operation with semi-synthetic medium with 50 and 100 g/l initial lactose concentrations are presented. The best result for beta-galactosidase...

  3. Decontamination of Ochratoxin-A Producing Aspergillus niger and Ochratoxin A in...

    OpenAIRE

    , N.G. Allam; , A. El-Raheem R. El-Shanshoury; , H.A. Emara; , A.Z. Zaky

    2016-01-01

    In this study twenty two medicinal and herbal plants among the common plants used in Egypt were collected and evaluated for fungal and ochratoxin contamination. Aspergillus, Penicillium, Fusarium, Botrytis and Cladosporium species were the most common fungi with different levels in most medicinal and food herbs under study. Out of the tested plants and the isolated fungi, black pepper contaminated with the highest amount of ochratoxin A and A. niger produced the highest amounts of ochratoxin ...

  4. Glucoamylases production of Aspergillus niger in solid state fermentation using a continuous counter-current reactor

    OpenAIRE

    Varzakas, T. H.; Roussos, Sevastianos; Arvanitoyannis, I. S.

    2008-01-01

    This work presents the continuous production of fungal biomass and glucoamylase by solid state fermentation (SSF) in a counter-current reactor adapted for this purpose. Pre-germinated conidia of Aspergillus niger were used as an inoculum, and sugarcane bagasse, embedded with a nutritive solution, was the solid support. The solids residence time distribution (RTD) was carried out by feeding one impule of blue-coloured humidified bagasse and its RTD was fixed at 20 h. This study demonstrated th...

  5. Fate and Role of Ammonium Ions during Fermentation of Citric Acid by Aspergillus niger

    OpenAIRE

    Papagianni, Maria; Wayman, Frank; Mattey, Michael

    2005-01-01

    Stoichiometric modeling of the early stages of the citric acid fermentation process by Aspergillus niger revealed that ammonium ions combine with a carbon-containing metabolite inside the cell, in a ratio 1:1, to form a nitrogen compound which is then excreted by the mycelium. High-performance liquid chromatography analysis identified glucosamine as the product of the relationship between glucose and ammonium during the early stages of the citric acid fermentation process. Slightly acidic int...

  6. Citric Acid Production by Aspergillus niger Cultivated on Parkia biglobosa Fruit Pulp

    OpenAIRE

    Auta, Helen Shnada; Abidoye, Khadijat Toyin; Tahir, Hauwa; Ibrahim, Aliyu Dabai; Aransiola, Sesan Abiodun

    2014-01-01

    The study was conducted to investigate the potential of Parkia biglobosa fruit pulp as substrate for citric acid production by Aspergillus niger. Reducing sugar was estimated by 3,5-dinitrosalicylic acid and citric acid was estimated spectrophotometrically using pyridine-acetic anhydride methods. The studies revealed that production parameters (pH, inoculum size, substrate concentration, incubation temperature, and fermentation period) had profound effect on the amount of citric acid produced...

  7. Attempts at improving citric acid fermentation by Aspergillus niger in beet-molasses medium.

    Science.gov (United States)

    Adham, Nehad Z

    2002-08-01

    Natural oils with high unsaturated fatty acids content when added at concentrations of 2% and 4% (v/v) to beet molasses (BM) medium caused a considerable increase in citric acid yield from Aspergillus niger. The fermentation capacities were also examined for production of citric acid using BM-oil media under different fermentation conditions. Maximum citric acid yield was achieved in surface culture in the presence of 4% olive oil after 12 days incubation.

  8. The changes in enzyme activities during citric acid fermentation by Aspergillus niger in solid culture.

    OpenAIRE

    桐村, 光太郎; 熊谷, 和夫; 森貞, 信也; "河邊, 誠一郎"; 宇佐美, 昭次; KOTARO, KIRIMURA; Kazuo, KUMAGAI; SHINYA, MORISADA; Seiichiro", "Kawabe; Shoji, Usami; 住友化学工業株式会社; 岡山理科大学基礎理学科; 早稲田大学理工学部応用化学科; Department of Applied Chemistry, Faculty of Science and Engineering, Kinki University

    1984-01-01

    The changes of enzyme activities during citric acid fermentation in solid culture were studied with a high citric acidproducing strain, Aspergillus niger Yang no. 2,cultivated in synthetic medium containing 2 g/l of gelatin and glass wool as carrier.With fermentation time, the specific activity ratio of fructose-bisphosphate aldolase to glucose-6-phosphate dehydrogenase increased. Of the TCA cycle enzmes, the specific activities of citrate synthetase and aconitate hydratase remained almost co...

  9. Control of Aspergillus niger with garlic, onion and leek extracts

    African Journals Online (AJOL)

    Administrator

    2007-02-19

    Feb 19, 2007 ... Condiments Chemistry,Microbiology ,Technology.Academic Press,. San Fransisco,pp 35-305. Rasooli I, Abyaneh MR (2004).Inhibitory effects of thyme oils on growth and aflatoxin production by Aspergillus parasiticus. Food Control. 15: 479-483. Rivlin RS (2001). Historical perspective on the use of garlic.

  10. Pectinolytic complex production by Aspergillus niger URM 4645 ...

    African Journals Online (AJOL)

    SAM

    2014-08-06

    Aug 6, 2014 ... ... Yadav PK, Yadav D, Yadav KDS (2009). Purification and characterization of pectin lyase produced by Aspergillus terricola and its application in retting of natural fibers. Appl. Biochem. Biotechnol. 159: 270-283. Zenebon O, Pascuet NS (2005). Métodos físico-químicos para análise de alimentos (4th ed.).

  11. Physiological characterisation of acuB deletion in Aspergillus niger

    DEFF Research Database (Denmark)

    Meijer, Susan Lisette; De Jongh, Willem Adriaan; Olsson, Lisbeth

    2009-01-01

    that a basal level of FacB activity exists under glucose-repressive conditions. In the present study, the effect of deletion of acuB on the physiology of A. niger was assessed. Differences in organic acid and acetate production, enzyme activities and extracellular amino and non-amino organic acid production...... were determined under glucose-repressing and -derepressing conditions. Furthermore, consumption of alternative carbon sources (e.g. xylose, citrate, lactate and succinate) was investigated. It was shown that AcuB has pleiotropic effects on the physiology of A. niger. The results indicate that metabolic...... pathways that are not directly involved in acetate metabolism are influenced by acuB deletion. Clear differences in organic acid consumption and production were detected between the a dagger acuB and reference strain. However, the hypothesis that AcuB is responsible for basal AcuA activity necessary...

  12. The effect of Aspergillus niger mutagenization on citric acid biosynthesis

    Directory of Open Access Journals (Sweden)

    Stanisław Walisch

    2014-08-01

    Full Text Available The industrial A. niger strain producing citric acid was mutagenized with the use of new chemical mutagens: free nitroxyl radicals. Strains of higher citric acid production yield were obtained. Citric acid was produced in a shorter time compared to the initial strain. During 6-12 months of storage most of the strains preserved their positive features which proves that mutants with profitable biotechnological properties were obtained. These mutants are used in industrial process.

  13. Manipueira como substrato na biossíntese de ácido cítrico por Aspergillus niger Citric acid production by Aspergillus niger from "manipueira", a manioc liquid residue

    Directory of Open Access Journals (Sweden)

    M. Leonel

    1995-08-01

    Full Text Available O trabalho teve por objetivo avaliar a viabilidade do uso da manipueira, residuo líquido resultante da prensagem da massa ralada de mandioca, como substrato na biossíntese de ácido cítrico por Aspergillus niger. Os meios de manipueira foram comparados nas mesmas condições de temperatura, a meios sintéticos, utilizados tradicionalmente. Em se tratando de proposta de um novo substrato, foi estudado o armazenamento do resíduo a temperatura ambiente por 72 horas, e realizada a caracterização físico-química da manipueira e dos meios elaborados com esse substrato. Foi avaliada a produção de ácido cítrico nos meios sintéticos e de manipueira. Verificou-se que a produção de ácido cítrico não diferiu quanto ao meio. Não foi observado crescimento do microrganismo nos meios de manipueira com concentrações acima de 70 mg/l de cianeto. Os resultados obtidos mostraram necessidade de maiores estudos para viabilizar o uso da manipueira como substrato na biossíntese de ácido cítrico por A. niger, principalmente no que diz respeito à liberação enzimática do cianeto.This research was made to evaluate the manipueira as substrat for citric acid biosynthesis by Aspergillus niger. The manipueira medium was compared in the same conditions of the synthetical medium, traditionally used in citric acid biosynthesis. The residue was kept at room temperature during 72 hours, and the physico-chemical characterization was made. The citric acid production did not change in the different media and the fungus did not grow hi the manipueira medium with concentrations about of 70 mg/l of cyanide. The results showed that more research is need to make manipueira utilization possible as a substrat for citric acid biosynthesis by Aspergillus niger.

  14. Molecular Cloning and Characteristic Features of a Novel Extracellular Tyrosinase from Aspergillus niger PA2.

    Science.gov (United States)

    Agarwal, Pragati; Singh, Jyoti; Singh, R P

    2017-05-01

    Aspergillus niger PA2, a novel strain isolated from waste effluents of food industry, is a potential extracellular tyrosinase producer. Enzyme activity and L-DOPA production were maximum when glucose and peptone were employed as C source and nitrogen source respectively in the medium and enhanced notably when the copper was supplemented, thus depicting the significance of copper in tyrosinase activity. Tyrosinase-encoding gene from the fungus was cloned, and amplification of the tyrosinase gene yielded a 1127-bp DNA fragment and 374 amino acid residue long product that encoded for a predicted protein of 42.3 kDa with an isoelectric point of 4.8. Primary sequence analysis of A. niger PA2 tyrosinase had shown that it had approximately 99% identity with that of A. niger CBS 513.88, which was further confirmed by phylogenetic analysis. The inferred amino acid sequence of A. niger tyrosinase contained two putative copper-binding sites comprising of six histidines, a characteristic feature for type-3 copper proteins, which were highly conserved in all tyrosinases throughout the Aspergillus species. When superimposed onto the tertiary structure of A. oryzae tyrosinase, the conserved residues from both the organisms occupied same spatial positions to provide a di-copper-binding peptide groove.

  15. Germination of Aspergillus niger conidia is triggered by nitrogen compounds related to L-amino acids.

    Science.gov (United States)

    Hayer, Kimran; Stratford, Malcolm; Archer, David B

    2014-10-01

    Conidial germination is fundamentally important to the growth and dissemination of most fungi. It has been previously shown (K. Hayer, M. Stratford, and D. B. Archer, Appl. Environ. Microbiol. 79:6924-6931, 2013, http://dx.doi.org/10.1128/AEM.02061-13), using sugar analogs, that germination is a 2-stage process involving triggering of germination and then nutrient uptake for hyphal outgrowth. In the present study, we tested this 2-stage germination process using a series of nitrogen-containing compounds for the ability to trigger the breaking of dormancy of Aspergillus niger conidia and then to support the formation of hyphae by acting as nitrogen sources. Triggering and germination were also compared between A. niger and Aspergillus nidulans using 2-deoxy-D-glucose (trigger), D-galactose (nontrigger in A. niger but trigger in A. nidulans), and an N source (required in A. niger but not in A. nidulans). Although most of the nitrogen compounds studied served as nitrogen sources for growth, only some nitrogen compounds could trigger germination of A. niger conidia, and all were related to L-amino acids. Using L-amino acid analogs without either the amine or the carboxylic acid group revealed that both the amine and carboxylic acid groups were essential for an L-amino acid to serve as a trigger molecule. Generally, conidia were able to sense and recognize nitrogen compounds that fitted into a specific size range. There was no evidence of uptake of either triggering or nontriggering compounds over the first 90 min of A. niger conidial germination, suggesting that the germination trigger sensors are not located within the spore. Copyright © 2014, American Society for Microbiology. All Rights Reserved.

  16. Mechanism for mercury tolerance in fungi. [Aspergillus niger; Rhizoctonia solani; Pythium ultimum

    Energy Technology Data Exchange (ETDEWEB)

    Ashworth, L.J. Jr.; Amin, J.V.

    1964-01-01

    Aspergillus niger was protected from mercury (Hg) poisoning by sulfhydryl (SH) compounds (glutathione and cysteine) in culture experiments, whereas Rhizoctonia solani and Pythium ultimum were not protected. Two- to 30-day-old mycelium of A. niger was found to be more tolerant to Hg than either younger or older mycelia. Mycelia of the other fungi were sensitive regardless of age. Mercury tolerance of A. niger mycelium was altered when sulfur nutrition was modified; it was greater after growth on a substrate containing reduced sulfur than after growth on one containing sulfate. The Hg tolerance of A. niger mycelium appears to be due to a pool of intracellular SH that is free of protein and that protects enzyme systems by forming complexes with Hg as it is taken up by the thallus. Potato-dextrose broth-grown mats of A. niger contained about 100 ..mu..g of SH/g of dry mycelia after 36 hr and after 5 days. R.. solani had about 9 ..mu..g of SH/g of mycelia after 5 days; a measurable amount was not recovered from P. ultimum. 20 references, 1 figure, 2 tables.

  17. HisB as novel selection marker for gene targeting approaches in Aspergillus niger.

    Science.gov (United States)

    Fiedler, Markus R M; Gensheimer, Tarek; Kubisch, Christin; Meyer, Vera

    2017-03-08

    For Aspergillus niger, a broad set of auxotrophic and dominant resistance markers is available. However, only few offer targeted modification of a gene of interest into or at a genomic locus of choice, which hampers functional genomics studies. We thus aimed to extend the available set by generating a histidine auxotrophic strain with a characterized hisB locus for targeted gene integration and deletion in A. niger. A histidine-auxotrophic strain was established via disruption of the A. niger hisB gene by using the counterselectable pyrG marker. After curing, a hisB - , pyrG - strain was obtained, which served as recipient strain for further studies. We show here that both hisB orthologs from A. nidulans and A. niger can be used to reestablish histidine prototrophy in this recipient strain. Whereas the hisB gene from A. nidulans was suitable for efficient gene targeting at different loci in A. niger, the hisB gene from A. niger allowed efficient integration of a Tet-on driven luciferase reporter construct at the endogenous non-functional hisB locus. Subsequent analysis of the luciferase activity revealed that the hisB locus is tight under non-inducing conditions and allows even higher luciferase expression levels compared to the pyrG integration locus. Taken together, we provide here an alternative selection marker for A. niger, hisB, which allows efficient homologous integration rates as well as high expression levels which compare favorably to the well-established pyrG selection marker.

  18. Physicochemical Properties Analysis and Secretome of Aspergillus niger in Fermented Rapeseed Meal.

    Science.gov (United States)

    Shi, Changyou; He, Jun; Yu, Jie; Yu, Bing; Mao, Xiangbing; Zheng, Ping; Huang, Zhiqing; Chen, Daiwen

    2016-01-01

    The nutrient digestibility and feeding value of rapeseed meal (RSM) for non-ruminant animals is poor due to the presence of anti-nutritional substances such as glucosinolate, phytic acid, crude fiber etc. In the present study, a solid state fermentation (SSF) using Aspergillus niger was carried out with the purpose of improving the nutritional quality of RSM. The chemical composition and physicochemical properties of RSM before and after fermentation were compared. To further understand possible mechanism of solid state fermentation, the composition of extracellular enzymes secreted by Aspergillus niger during fermentation was analysed using two-dimentional difference gel electrophoresis (2D-DIGE) combined with matrix assisted laser desorption ionization-time of flight-mass spectrometer (MALDI-TOF-MS). Results of the present study indicated that SSF had significant effects on chemical composition of RSM. The fermented rapeseed meal (FRSM) contained more crude protein (CP) and amino acid (AA) (except His) than unfermented RSM. Notably, the small peptide in FRSM was 2.26 time larger than that in unfermented RSM. Concentrations of anti-nutritional substrates in FRSM including neutral detergent fiber (NDF), glucosinolates, isothiocyanate, oxazolidithione, and phytic acid declined (P niger fermentation disrupted the surface structure, changed macromolecular organic compounds, and reduced the protein molecular weights of RSM substrate. Total proteins of raw RSM and FRSM were separated and 51 protein spots were selected for mass spectrometry according to 2D-DIGE map. In identified proteins, there were 15 extracellular hydrolases secreted by A. niger including glucoamylase, acid protease, beta-glucanase, arabinofuranosidase, xylanase, and phytase. Some antioxidant related enzymes also were identified. These findings suggested that A. niger is able to secrete many extracellular degradation enzymes (especially lignocellulosic hydrolyzing enzymes, acid proteases and phytase

  19. Responses of Aspergillus niger to selected environmental factors

    Directory of Open Access Journals (Sweden)

    Šimonovičová Alexandra

    2017-12-01

    Full Text Available Four wild type strains of A. niger were collected from soil and stream sediments representing environments with variable level of As, Sb, Al, Fe, Cd, Cu, and Zn contamination. Banská Štiavnica-Šobov (S, Pezinok-Kolársky vrch (P and Slovinky (Sl represent contaminated localities. Locality Gabčíkovo (G was as a control site. The influence of toxic elements in these substrates on fungal growth, colony size, enzymatic activity, production of organic acids and their pelletization in water suspensions with montmorillonite was studied. The aim of our study was to find out how the wild type strains from (contaminated environment will behave in different model solutions. We also wanted to add some new information in this area of study, because that there is some gap in the available knowledge.

  20. Optimization of ellagitannase production by Aspergillus niger GH1 by solid-state fermentation.

    Science.gov (United States)

    de la Cruz, Reynaldo; Ascacio, Juan A; Buenrostro, Juan; Sepúlveda, Leonardo; Rodríguez, Raúl; Prado-Barragán, Arely; Contreras, Juan C; Aguilera, Antonio; Aguilar, Cristóbal N

    2015-01-01

    Ellagic acid is one of the most bioactive antioxidants with important applications in pharmaceutical, cosmetic, and food industries. However, there are few biotechnological processes developed for its production, because it requires precursors (ellagitannins) and the corresponding biocatalyst (ellagitannase). The aim of this study was to optimize the culture conditions for ellagitannase production by Aspergillus niger in solid-state fermentation (SSF). The bioprocess was carried out into a column bioreactor packed with polyurethane foam impregnated with an ellagitannins solution as carbon source. Four strains of Aspergillus niger (PSH, GH1, HT4, and HC2) were evaluated for ellagitannase production. The study was performed in two experimental steps. A Plackett-Burman design was used to determine the influencing parameters on ellagitannase production. Ellagitannins concentration, KCl, and MgSO4 were determined to be the most significant parameters. Box-Behnken design was used to define the interaction of the selected parameters. The highest enzyme value was obtained by A. niger PSH at concentrations of 7.5 g/L ellagitannins, 3.04 g/L KCl, and 0.76 g/L MgSO4. The methodology followed here allowed increasing the ellagitannase activity 10 times over other researcher results (938.8 U/g ellagitannins). These results are significantly higher than those reported previously and represent an important contribution for the establishment of a new bioprocess for ellagic acid and ellagitannase production.

  1. Comparative Secretome Analysis of Aspergillus niger, Trichoderma reesei, and Penicillium oxalicum During Solid-State Fermentation.

    Science.gov (United States)

    Gong, Weili; Zhang, Huaiqiang; Liu, Shijia; Zhang, Lili; Gao, Peiji; Chen, Guanjun; Wang, Lushan

    2015-11-01

    Filamentous fungi such as Aspergillus spp., Trichoderma spp., and Penicillium spp. are frequently used to produce high concentrations of lignocellulosic enzymes. This study examined the discrepancies in the compositions and dynamic changes in the extracellular enzyme systems secreted by Aspergillus niger ATCC1015, Trichoderma reesei QM9414, and Penicillium oxalicum 114-2 cultured on corn stover and wheat bran. The results revealed different types and an abundance of monosaccharides and oligosaccharides were released during incubation, which induced the secretion of diverse glycoside hydrolases. Both the enzyme activities and isozyme numbers of the three fungal strains increased with time. A total of 279, 161, and 183 secretory proteins were detected in A. niger, T. reesei, and P. oxalicum secretomes, respectively. In the A. niger secretomes, more enzymes involved in the degradation of (galacto)mannan, xyloglucan, and the backbone of pectin distributed mostly in dicots were detected. In comparison, although P. oxalicum 114-2 hardly secreted any xyloglucanases, the diversities of enzymes involved in the degradation of xylan and β-(1,3;1,4)-D-glucan commonly found in monocots were higher. The cellulase system of P. oxalicum 114-2 was more balanced. The degradation preference provided a new perspective regarding the recomposition of lignocellulosic enzymes based on substrate types.

  2. Mutualistic interaction between Salmonella enterica and Aspergillus niger and its effects on Zea mays colonization.

    Science.gov (United States)

    Balbontín, Roberto; Vlamakis, Hera; Kolter, Roberto

    2014-11-01

    Salmonella Typhimurium inhabits a variety of environments and is able to infect a broad range of hosts. Throughout its life cycle, some hosts can act as intermediates in the path to the infection of others. Aspergillus niger is a ubiquitous fungus that can often be found in soil or associated to plants and microbial consortia. Recently, S. Typhimurium was shown to establish biofilms on the hyphae of A. niger. In this work, we have found that this interaction is stable for weeks without a noticeable negative effect on either organism. Indeed, bacterial growth is promoted upon the establishment of the interaction. Moreover, bacterial biofilms protect the fungus from external insults such as the effects of the anti-fungal agent cycloheximide. Thus, the Salmonella-Aspergillus interaction can be defined as mutualistic. A tripartite gnotobiotic system involving the bacterium, the fungus and a plant revealed that co-colonization has a greater negative effect on plant growth than colonization by either organism in dividually. Strikingly, co-colonization also causes a reduction in plant invasion by S. Typhimurium. This work demonstrates that S. Typhimurium and A. niger establish a mutualistic interaction that alters bacterial colonization of plants and affects plant physiology. © 2014 The Authors. Microbial Biotechnology published by John Wiley & Sons Ltd and Society for Applied Microbiology.

  3. Polygalacturonase gene pgxB in Aspergillus niger is a virulence factor in apple fruit.

    Science.gov (United States)

    Liu, Cheng-Qian; Hu, Kang-Di; Li, Ting-Ting; Yang, Ying; Yang, Feng; Li, Yan-Hong; Liu, He-Ping; Chen, Xiao-Yan; Zhang, Hua

    2017-01-01

    Aspergillus niger, a saprophytic fungus, is widely distributed in soil, air and cereals, and can cause postharvest diseases in fruit. Polygalacturonase (PG) is one of the main enzymes in fungal pathogens to degrade plant cell wall. To evaluate whether the deletion of an exo-polygalacturonase gene pgxB would influence fungal pathogenicity to fruit, pgxB gene was deleted in Aspergillus niger MA 70.15 (wild type) via homologous recombination. The ΔpgxB mutant showed similar growth behavior compared with the wild type. Pectin medium induced significant higher expression of all pectinase genes in both wild type and ΔpgxB in comparison to potato dextrose agar medium. However, the ΔpgxB mutant was less virulent on apple fruits as the necrosis diameter caused by ΔpgxB mutant was significantly smaller than that of wild type. Results of quantitive-PCR showed that, in the process of infection in apple fruit, gene expressions of polygalacturonase genes pgaI, pgaII, pgaA, pgaC, pgaD and pgaE were enhanced in ΔpgxB mutant in comparison to wild type. These results prove that, despite the increased gene expression of other polygalacturonase genes in ΔpgxB mutant, the lack of pgxB gene significantly reduced the virulence of A. niger on apple fruit, suggesting that pgxB plays an important role in the infection process on the apple fruit.

  4. Degradation of chlorimuron-ethyl by Aspergillus niger isolated from agricultural soil.

    Science.gov (United States)

    Sharma, Seema; Banerjee, Kaushik; Choudhury, Partha P

    2012-12-01

    Chlorimuron-ethyl, ethyl-2-[[[[(4-methoxy-6-chloro-pyrimidin-2-yl)amino]carbonyl]amino] sulfonyl]benzoate, is used as a pre- and postemergence herbicide for the control of important broadleaved weeds in soybean and maize. Due to its phytotoxicity to rotation crops, concerns regarding chlorimuron contamination of soil and water have been raised. Although it is degraded in the agricultural environment primarily via pH- and temperature-dependent chemical hydrolysis, microbial transformation also has an important role. Fungi such as Fusarium and Alternaria are unable to survive in artificial media containing chlorimuron-ethyl at 25 mg L(-1) . However, Aspergillus niger survived in minimal broth containing chlorimuron at 2 mg mL(-1) . Aspergillus niger degraded the herbicide to harvest energy through two major routes of degradation. One route involves the cleavage of the sulfonylurea bridge, resulting in the formation of two major metabolites, namely ethyl-2-aminosulfonylbenzoate (I) and 4-methoxy-6-chloro-2-amino-pyrimidine (II). The other route is the cleavage of sulfonylamide linkage, which generates the metabolite N-(4-methoxy-6-chloropyrimidin-2-yl) urea (III). Two other metabolites, saccharin (IV) and N-methyl saccharin (V), formed from metabolite II, were also identified. A metabolic pathway for the degradation of chlorimuron-ethyl by A. niger has been proposed. © 2012 Federation of European Microbiological Societies. Published by Blackwell Publishing Ltd. All rights reserved.

  5. Formation of Sclerotia and Production of Indoloterpenes by Aspergillus niger and Other Species in Section Nigri

    Science.gov (United States)

    Frisvad, Jens C.; Petersen, Lene M.; Lyhne, E. Kirstine; Larsen, Thomas O.

    2014-01-01

    Several species in Aspergillus section Nigri have been reported to produce sclerotia on well-known growth media, such as Czapek yeast autolysate (CYA) agar, with sclerotia considered to be an important prerequisite for sexual development. However Aspergillus niger sensu stricto has not been reported to produce sclerotia, and is thought to be a purely asexual organism. Here we report, for the first time, the production of sclerotia by certain strains of Aspergillus niger when grown on CYA agar with raisins, or on other fruits or on rice. Up to 11 apolar indoloterpenes of the aflavinine type were detected by liquid chromatography and diode array and mass spectrometric detection where sclerotia were formed, including 10,23-dihydro-24,25-dehydroaflavinine. Sclerotium induction can thus be a way of inducing the production of new secondary metabolites from previously silent gene clusters. Cultivation of other species of the black aspergilli showed that raisins induced sclerotium formation by A. brasiliensis, A. floridensis A. ibericus, A. luchuensis, A. neoniger, A. trinidadensis and A. saccharolyticus for the first time. PMID:24736731

  6. Xylanases from Aspergillus niger, Aspergillus niveus and Aspergillus ochraceus produced under solid-state fermentation and their application in cellulose pulp bleaching.

    Science.gov (United States)

    Betini, J H A; Michelin, M; Peixoto-Nogueira, S C; Jorge, J A; Terenzi, H F; Polizeli, M L T M

    2009-10-01

    This study describes the production of xylanases from Aspergillus niveus, A. niger, and A. ochraceus under solid-state fermentation using agro-industrial residues as substrates. Enzyme production was improved using a mixture of wheat bran and yeast extract or peptone. When a mixture of corncob and wheat bran was used, xylanase production from A. niger and A. ochraceus increased by 18%. All cultures were incubated at 30 degrees C at 70-80% relative humidity for 96 h. For biobleaching assays, 10 or 35 U of xylanase/g dry cellulose pulp were incubated at pH 5.5 for 1 or 2 h, at 55 degrees C. The delignification efficiency was 20%, the brightness (percentage of ISO) increased two to three points and the viscosity was maintained confirming the absence of cellulolytic activity. These results indicated that the use of xylanases could help to reduce the amount of chlorine compounds used in cellulose pulp treatment.

  7. Removal of silver nanoparticles using live and heat shock Aspergillus niger cultures.

    Science.gov (United States)

    Gomaa, Ola M

    2014-06-01

    Silver nanoparticles (SNPs) are extensively used in many industrial and medical applications; however, the impact of their release in the environment is still considered an understudied field. In the present work, SNPs present in aqueous lab waste water (average size of 30 nm) were used to determine their impact on microflora if released in soil rhizosphere and sewage waste water. The results showed that 24 h incubation with different SNP concentrations resulted in a 2.6-fold decrease for soil rhizosphere microflora and 7.45-fold decrease for sewage waste water microflora, both at 24 ppm. Live and heat shock (50 and 70 °C) Aspergillus niger cultures were used to remove SNP waste, the results show 76.6, 81.74 and 90.8 % SNP removal, respectively after 3 h incubation. There was an increase in the log total bacterial count again after SNP removal by A. niger in the following order: live A. niger niger niger. The pH value decreased from 5.8 to 3.8 in the same order suggesting the production of an acid in the culture media. Scanning electron microscopy images showed agglomeration and/or complexation of SNP particles, in a micron size, in between the fungal mycelia, hence settling on and in between the mycelial network. The results suggest that silver was reduced again and agglomerated and/or chelated together in its oxidized form by an acid in A. niger media. More studies are recommended to determine the acid and the heat shock proteins to confirm the exact mode of action.

  8. In Vitro Efficacy of Continuous Mild High Temperature on the Biofilm Formation of Aspergillus Niger.

    Science.gov (United States)

    Zeng, Rong; Tong, Jian Bo; Liu, Yu Zhen; Chen, Qing; Lin, Tong; Li, Min; Lü, Gui Xia

    2017-12-20

    Objective To investigate whether continuous mild high temperature (increased temperature without causing significant damage to host cells) can inhibit the biofilm formation of Aspergillus niger (A.niger) and its vitality.Methods A.niger biofilms were formed on a coverslip in 24-well tissue culture plate and were checked at the time points 4,8,10,16,24,48 and 72 hours.Confocal laser scanning microscopy (CLSM) was used to image and quantify A.niger biofilm formation under three different continuous mild high temperatures at 37℃,39℃,and 41℃.Furthermore,2,3-bis(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide (XTT) assay was used to quantify the dynamic growth of A.niger biofilm under the above conditions.Results Compared with the culture condition 37℃,CLSM analysis at 39℃ or 41℃ showed that higher temperature induced later germination at 4 hours (t=8.603,P=0.047;t=14.550,P=0.008),poorer hyphal elongation at 8 hours(t=35.118,P=0.039;t=63.450,P=0.006),poorer polar growth,and reduced biofilm thickness from 10 to 24 hours.The XTT assay showed that higher temperature (39℃ or 41℃) lead to lower vitality at 10 hours,higher vitality at 16 hours,but finally lower vitality from 24 to 72 hours (t=24.262,P=0.038;t=7.556,P=0.031).Conclusion Continuous mild high temperature may have a negative regulatory effect on biofilm formation of A.niger and its vitality.

  9. Adsorção de índigo carmim em biomassas mortas de aspergillus niger Adsorption of indigo carmine on dead biomass of aspergillus niger

    Directory of Open Access Journals (Sweden)

    Karina Oliveira Chaves

    2008-12-01

    Full Text Available As indústrias têxteis geram efluentes ricos em corantes durante toda a sua cadeia produtiva. Neste estudo, ensaios cinéticos foram realizados utilizando biomassas mortas de Aspergillus niger, diferentemente tratadas, como adsorvente para a remoção de índigo carmim a partir de soluções aquosas. Ensaio cinético com carvão ativado comercial também foi realizado para comparação. Os resultados obtidos indicam que a biomassa fúngica sem tratamento algum foi capaz de remover 95% do índigo carmim da solução aquosa, remoção equivalente àquela obtida pelo carvão ativado comercial. Os experimentos mostraram que os tratamentos ácidos melhoraram consideravelmente a cinética da remoção do corante, não influenciando, contudo, a capacidade adsortiva máxima do material. Os tratamentos salinos e alcalinos diminuíram tanto a capacidade máxima quanto a cinética de adsorção.Textile industries produce effluents rich in dyes throughout its production chain. In this study, kinetic experiments were carried using the dead and treated biomass of Aspergillus niger for indigo carmine removal from the synthetic aqueous solutions of this dye. A kinetic run with activated carbon was also carried for comparison. The obtained results suggest that non-treated fungal biomass has been capable of remove 95% of carmine indigo from the aqueous solutions. This removal was similar to that obtained with activated carbon. The results also indicate that acid treatments considerably improve the kinetic of the dye removal, but they are not influenced the maximum adsorptive capacity of the adsorbent. Moreover, saline and alkaline treatments decreased both the maximum capacity and the kinetic of adsorption.

  10. Lipase of Aspergillus niger NCIM 1207: A Potential Biocatalyst for Synthesis of Isoamyl Acetate.

    Science.gov (United States)

    Mhetras, Nutan; Patil, Sonal; Gokhale, Digambar

    2010-10-01

    Commercial lipase preparations and mycelium bound lipase from Aspergillus niger NCIM 1207 were used for esterification of acetic acid with isoamyl alcohol to obtain isoamyl acetate. The esterification reaction was carried out at 30°C in n-hexane with shaking at 120 rpm. Initial reaction rates, conversion efficiency and isoamyl acetate concentration obtained using Novozyme 435 were the highest. Mycelium bound lipase of A. niger NCIM 1207 produced maximal isoamyl acetate formation at an alcohol/acid ratio of 1.6. Acetic acid at higher concentrations than required for the critical alcohol/acid ratio lower than 1.3 and higher than 1.6 resulted in decreased yields of isoamyl acetate probably owing to lowering of micro-aqueous environmental pH around the enzyme leading to inhibition of enzyme activity. Mycelium bound A. niger lipase produced 80 g/l of isoamyl acetate within 96 h even though extremely less amount of enzyme activity was used for esterification. The presence of sodium sulphate during esterification reaction at higher substrate concentration resulted in increased conversion efficiency when we used mycelium bound enzyme preparations of A. niger NCIM 1207. This could be due to removal of excess water released during esterification reaction by sodium sulphate. High ester concentration (286.5 g/l) and conversion (73.5%) were obtained within 24 h using Novozyme 435 under these conditions.

  11. An inducible tool for random mutagenesis in Aspergillus niger based on the transposon Vader.

    Science.gov (United States)

    Paun, Linda; Nitsche, Benjamin; Homan, Tim; Ram, Arthur F; Kempken, Frank

    2016-07-01

    The ascomycete Aspergillus niger is widely used in the biotechnology, for instance in producing most of the world's citric acid. It is also known as a major food and feed contaminant. While generation of gene knockouts for functional genomics has become feasible in ku70 mutants, analyzing gene functions or metabolic pathways remains a laborious task. An unbiased transposon-based mutagenesis approach may aid this process of analyzing gene functions by providing mutant libraries in a short time. The Vader transposon is a non-autonomous DNA-transposon, which is activated by the homologous tan1-transposase. However, in the most commonly used lab strain of A. niger (N400 strain and derivatives), we found that the transposase, encoded by the tan1 gene, is mutated and inactive. To establish a Vader transposon-based mutagenesis system in the N400 background, we expressed the functional transposase of A. niger strain CBS 513.88 under the control of an inducible promoter based on the Tet-on system, which is activated in the presence of the antibiotic doxycycline (DOX). Increasing amounts of doxycycline lead to higher Vader excision frequencies, whereas little to none activity of Vader was observed without addition of doxycycline. Hence, this system appears to be suitable for producing stable mutants in the A. niger N400 background.

  12. Type III polyketide synthase is involved in the biosynthesis of protocatechuic acid in Aspergillus niger.

    Science.gov (United States)

    Lv, Yangyong; Xiao, Jing; Pan, Li

    2014-11-01

    Genomic studies have shown that not only plants but also filamentous fungi contain type III polyketide synthases. To study the function of type III polyketide synthase (AnPKSIII) in Aspergillus niger, a deletion strain (delAnPKSIII) and an overexpression strain (oeAnPKSIII) were constructed in A. niger MA169.4, a derivative of the wild-type (WT) A. niger ATCC 9029 that produces large quantities of gluconic acid. Alterations in the metabolites were analyzed by HPLC when the extract of the overexpression strain was compared with extracts of the WT and deletion strains. Protocatechuic acid (PCA; 3,4-dihydroxybenzoic acid, 3.2 mg/l) was isolated and identified as the main product of AnPKSIII when inductively expressed in A. niger MA169.4. The molecular weight of PCA was 154.1 (m/z 153.1 [M-H](-)), was detected by ESI-MS in the negative ionization mode, and (1)H and (13)C NMR data confirmed its structure.

  13. Simultaneous Production of Amyloglucosidase and Exo-Polygalacturonase by Aspergillus niger in a Rotating Drum Reactor.

    Science.gov (United States)

    Colla, Eliane; Santos, Lucielen Oliveira; Deamici, Kricelle; Magagnin, Glênio; Vendruscolo, Mauricio; Costa, Jorge Alberto Vieira

    2017-02-01

    Simultaneous production of amyloglucosidase (AMG) and exo-polygalacturonase (exo-PG) was carried out by Aspergillus niger in substrate of defatted rice bran in a rotating drum bioreactor (RDB) and studied by a 3(1) × 2(2) factorial experimental design. Variables under study were A. niger strains (A. niger NRRL 3122 and A. niger t0005/007-2), types of inoculum (spore suspension and fermented bran), and types of inducer (starch, pectin, and a mix of both). Solid-state fermentation process (SSF) was conducted at 30 °C under 60-vvm aeration for 96 h in a pilot scale. Production of AMG and exo-PG was significantly affected by the fungal strain and the type of inoculum, but inducers did not trigger any significant effect, an evidence of the fact that these enzymes are constitutive. The maximum activity of exo-PG was 84 U gdm(-1) whereas the maximum yield of AMG was 886.25 U gdm(-1).

  14. Salmonella biofilm formation on Aspergillus niger involves cellulose--chitin interactions.

    Directory of Open Access Journals (Sweden)

    Maria T Brandl

    Full Text Available Salmonella cycles between host and nonhost environments, where it can become an active member of complex microbial communities. The role of fungi in the environmental adaptation of enteric pathogens remains relatively unexplored. We have discovered that S. enterica Typhimurium rapidly attaches to and forms biofilms on the hyphae of the common fungus, Aspergillus niger. Several Salmonella enterica serovars displayed a similar interaction, whereas other bacterial species were unable to bind to the fungus. Bacterial attachment to chitin, a major constituent of fungal cell walls, mirrored this specificity. Pre-incubation of S. Typhimurium with N-acetylglucosamine, the monomeric component of chitin, reduced binding to chitin beads by as much as 727-fold and inhibited attachment to A. niger hyphae considerably. A cellulose-deficient mutant of S. Typhimurium failed to attach to chitin beads and to the fungus. Complementation of this mutant with the cellulose operon restored binding to chitin beads to 79% of that of the parental strain and allowed for attachment and biofilm formation on A. niger, indicating that cellulose is involved in bacterial attachment to the fungus via the chitin component of its cell wall. In contrast to cellulose, S. Typhimurium curli fimbriae were not required for attachment and biofilm development on the hyphae but were critical for its stability. Our results suggest that cellulose-chitin interactions are required for the production of mixed Salmonella-A. niger biofilms, and support the hypothesis that encounters with chitinaceous alternate hosts may contribute to the ecological success of human pathogens.

  15. Risk assessment of fungal spoilage: A case study of Aspergillus niger on yogurt.

    Science.gov (United States)

    Gougouli, Maria; Koutsoumanis, Konstantinos P

    2017-08-01

    A quantitative risk assessment model of yogurt spoilage by Aspergillus niger was developed based on a stochastic modeling approach for mycelium growth by taking into account the important sources of variability such as time-temperature conditions during the different stages of chill chain and individual spore behavior. Input parameters were fitted to the appropriate distributions and A. niger colony's diameter at each stage of the chill chain was estimated using Monte Carlo simulation. By combining the output of the growth model with the fungus prevalence, that can be estimated by the industry using challenge tests, the risk of spoilage translated to number of yogurt cups in which a visible mycelium of A. niger is being formed at the time of consumption was assessed. The risk assessment output showed that for a batch of 100,000 cups in which the percentage of contaminated cups with A. niger was 1% the predicted numbers (median (5(th), 95(th) percentiles)) of the cups with a visible mycelium at consumption time were 8 (5, 14). For higher percentages of 3, 5 and 10 the predicted numbers (median (5(th), 95(th) percentiles)) of the spoiled cups at consumption time were estimated to be 24 (16, 35), 39 (29, 52) and 80 (64, 94), respectively. The developed model can lead to a more effective risk-based quality management of yogurt and support the decision making in yogurt production. Copyright © 2017 Elsevier Ltd. All rights reserved.

  16. Air pressure pulsation solid state fermentation of feruloyl esterase by Aspergillus niger.

    Science.gov (United States)

    Zeng, W; Chen, H Z

    2009-02-01

    Air pressure pulsation solid state fermentation (APP-SSF) was applied to produce feruloyl esterase (FAE) by Aspergillus niger. With the optimization of some variables by orthogonal design, the optimal condition obtained was 0.2 MPa (gauge pressure) of high pressure intensity, 30 min of low pressure duration and 20s of high pressure duration. Based on the optimized condition, the APP-SSF achieved the reasonable enzyme yield of 881 mU/g at 48 h, which was 58% more than that by static solid state fermentation (static SSF) at 72 h. By comparison of two fermentation methods in temperature, O(2) and CO(2) concentration, and respiration intensity, it was concluded that APP-SSF enhanced heat and mass transfer of fermentation system and strengthened the metabolism of microorganisms. The APP-SSF had a greatly positive effect on FAE production by A. niger, by enhancing mass and heat transfer and activating growth and metabolism.

  17. Morphological transitions under oxidative stress in response to metabolite formation in Aspergillus niger.

    Science.gov (United States)

    Lv, Yangyong; Zhou, Feng; Wang, Bin; Pan, Li

    2015-03-01

    Oxidative stress is associated with metabolite formation in fungi. In contrast to an Aspergillus niger wild-type strain, a sclerotia-formation regulator ansclR deletion strain demonstrated increased susceptibility to oxidative stress and reduced transcription of the catalase gene, catB, while an ansclR overexpression strain showed enhanced resistance to oxidative stress and increased expression of catB. In addition, ansclR complementation strain expressed a wild-type level of catB. The ansclR overexpression strain also produced the same metabolites as the wild type strain treated with H2O2. Furthermore, LC-MS, NMR, and IR analyses showed that the main metabolite was a steroid analog. Our study adds new clues to oxidative stress-related factors and metabolite formation in A. niger.

  18. Efficient Conversion of Inulin to Inulooligosaccharides through Endoinulinase from Aspergillus niger.

    Science.gov (United States)

    Xu, Yanbing; Zheng, Zhaojuan; Xu, Qianqian; Yong, Qiang; Ouyang, Jia

    2016-03-30

    Inulooligosaccharides (IOS) represent an important class of oligosaccharides at industrial scale. An efficient conversion of inulin to IOS through endoinulinase from Aspergillus niger is presented. A 1482 bp codon optimized gene fragment encoding endoinulinase from A. niger DSM 2466 was cloned into pPIC9K vector and was transformed into Pichia pastoris KM71. Maximum activity of the recombinant endoinulinase, 858 U/mL, was obtained at 120 h of the high cell density fermentation process. The optimal conditions for inulin hydrolysis using the recombinant endoinulinase were investigated. IOS were harvested with a high concentration of 365.1 g/L and high yield up to 91.3%. IOS with different degrees of polymerization (DP, mainly DP 3-6) were distributed in the final reaction products.

  19. The molecular and genetic basis of conidial pigmentation in Aspergillus niger

    DEFF Research Database (Denmark)

    Jørgensen, Thomas R.; Park, Joohae; Arentshorst, Mark

    2011-01-01

    A characteristic hallmark of Aspergillus niger is the formation of black conidiospores. We have identified four loci involved in spore pigmentation of A. niger by using a combined genomic and classical complementation approach. First, we characterized a newly isolated color mutant, colA, which...... lacked pigmentation resulting in white or colorless conidia. Pigmentation of the colA mutant was restored by a gene (An12g03950) which encodes a putative 4′phosphopantetheinyl transferase protein (PptA). 4′Phosphopantetheinyl transferase activity is required for the activation of Polyketide Synthases...... (PKSs) and/or Non-Ribosomal Peptide Synthases (NRPSs). The loci whose mutation resulted in fawn, olive, and brown color phenotypes were identified by complementation. The fawn phenotype was complemented by a PKS protein (FwnA, An09g05730), the ovlA mutant by An14g05350 (OlvA) and the brnA mutant by An14...

  20. Enhancement of citric acid production with ram horn hydrolysate by Aspergillus niger.

    Science.gov (United States)

    Kurbanoglu, Esabi Basaran

    2004-03-01

    The potential use of ram horn hydrolysate (RHH) as a supplement for improvement of citric acid production by Aspergillus niger NRRL 330 was studied. For this purpose, first RHH was produced. Ram horns were hydrolyzed by treating with acid (6 N-H2SO4) and the RHH was obtained. With the addition of RHH to the fermentation medium with a final concentration of 4% (optimal concentration), citric acid value reached a maximum value (94 g/l), which is 52% higher than that of the control experiment. The addition of 4% (v/v) RHH enhanced citric acid accumulation, reduced residual sugar concentration and stimulated mycelial growth. Adding 4% RHH had no adverse effects on A. niger. As a result, RHH was found to be suitable as a valuable supplement for citric acid production in the submerged fermentation.

  1. Isotherms, thermodynamic and mechanism studies of removal of low concentration uranium (VI) by Aspergillus niger.

    Science.gov (United States)

    Wang, Xiaoyu; Wang, Tieshan; Zheng, Xinyan; Shen, Yanghao; Lu, Xia

    2017-06-01

    In order to develop an effective and economical method for removing low concentration radioactive wastewater of uranium, the biomass of 'CMCC(F)-98003' Aspergillus niger was investigated in a batch system. The maximum uranium adsorption capacity of 12.5 mg g -1 was obtained at the initial uranium concentration of 0.75 mg L -1 . The biosorption data on a biomass concentration of 0.029 g L -1 fitted well to the Freundlich isotherm with a correlation coefficient (R 2 ) of 0.987. The calculated thermodynamic parameters showed that the biosorption of uranium ions was endothermic (ΔH° uranium precipitation were formed on the cell surfaces after biosorption, and the functional groups of -CH, N-H, -COOH, P = O and the carbohydrates and alcohols were involved in the biosorption process between A. niger and uranium ions.

  2. Review of secondary metabolites and mycotoxins from the Aspergillus niger group

    DEFF Research Database (Denmark)

    Nielsen, Kristian Fog; Mogensen, Jesper Mølgaard; Johansen, Maria

    2009-01-01

    Filamentous fungi in the Aspergillus section Nigri (the black aspergilli) represent some of the most widespread food and feed contaminants known but they are also some of the most important workhorses used by the biotechnological industry. The Nigri section consists of six commonly found species...... (excluding A. aculeatus and its close relatives) from which currently 145 different secondary metabolites have been isolated and/or detected. From a human and animal safety point of view, the mycotoxins ochratoxin A (from A. carbonarius and less frequently A. niger) and fumonisin B2 (from A. niger...... for monitoring in food, feed, and biotechnology products as well as for a better toxicological evaluation, since they are often produced in large amounts by the black aspergilli. For chemical differentiation/identification of the less toxic species the diketopiperazine asperazine can be used as a positive marker...

  3. Physicochemical Properties Analysis and Secretome of Aspergillus niger in Fermented Rapeseed Meal

    Science.gov (United States)

    Shi, Changyou; He, Jun; Yu, Jie; Yu, Bing; Mao, Xiangbing; Zheng, Ping; Huang, Zhiqing; Chen, Daiwen

    2016-01-01

    The nutrient digestibility and feeding value of rapeseed meal (RSM) for non-ruminant animals is poor due to the presence of anti-nutritional substances such as glucosinolate, phytic acid, crude fiber etc. In the present study, a solid state fermentation (SSF) using Aspergillus niger was carried out with the purpose of improving the nutritional quality of RSM. The chemical composition and physicochemical properties of RSM before and after fermentation were compared. To further understand possible mechanism of solid state fermentation, the composition of extracellular enzymes secreted by Aspergillus niger during fermentation was analysed using two-dimentional difference gel electrophoresis (2D-DIGE) combined with matrix assisted laser desorption ionization—time of flight—mass spectrometer (MALDI-TOF-MS). Results of the present study indicated that SSF had significant effects on chemical composition of RSM. The fermented rapeseed meal (FRSM) contained more crude protein (CP) and amino acid (AA) (except His) than unfermented RSM. Notably, the small peptide in FRSM was 2.26 time larger than that in unfermented RSM. Concentrations of anti-nutritional substrates in FRSM including neutral detergent fiber (NDF), glucosinolates, isothiocyanate, oxazolidithione, and phytic acid declined (P fermentation disrupted the surface structure, changed macromolecular organic compounds, and reduced the protein molecular weights of RSM substrate. Total proteins of raw RSM and FRSM were separated and 51 protein spots were selected for mass spectrometry according to 2D-DIGE map. In identified proteins, there were 15 extracellular hydrolases secreted by A. niger including glucoamylase, acid protease, beta-glucanase, arabinofuranosidase, xylanase, and phytase. Some antioxidant related enzymes also were identified. These findings suggested that A. niger is able to secrete many extracellular degradation enzymes (especially lignocellulosic hydrolyzing enzymes, acid proteases and phytase

  4. Aspergillus niger Enhance Bioactive Compounds Biosynthesis As Well As Expression of Functional Genes in Adventitious Roots of Glycyrrhiza uralensis Fisch.

    Science.gov (United States)

    Li, Jing; Wang, Juan; Li, Jinxin; Liu, Dahui; Li, Hongfa; Gao, Wenyuan; Li, Jianli; Liu, Shujie

    2016-02-01

    In the present study, the culture conditions for the accumulation of Glycyrrhiza uralensis adventitious root metabolites in balloon-type bubble bioreactors (BTBBs) have been optimized. The results of the culture showed that the best culture conditions were a cone angle of 90° bioreactor and 0.4-0.6-0.4-vvm aeration volume. Aspergillus niger can be used as a fungal elicitor to enhance the production of defense compounds in plants. With the addition of a fungal elicitor (derived from Aspergillus niger), the maximum accumulation of total flavonoids (16.12 mg g(-1)) and glycyrrhetinic acid (0.18 mg g(-1)) occurred at a dose of 400 mg L(-1) of Aspergillus niger resulting in a 3.47-fold and 1.8-fold increase over control roots. However, the highest concentration of polysaccharide (106.06 mg g(-1)) was achieved with a mixture of elicitors (Aspergillus niger and salicylic acid) added to the medium, resulting in a 1.09-fold increase over Aspergillus niger treatment alone. Electrospray ionization tandem mass spectrometry (ESI-MS(n)) analysis was performed, showing that seven compounds were present after treatment with the elicitors, including uralsaponin B, licorice saponin B2, liquiritin, and (3R)-vestitol, only identified in the mixed elicitor treatment group. It has also been found that elicitors (Aspergillus niger and salicylic acid) significantly upregulated the expression of the cinnamate 4-hydroxylase (C4H), β-amyrin synthase (β-AS), squalene epoxidase (SE) and a cytochrome P450 monooxygenase (CYP72A154) genes, which are involved in the biosynthesis of bioactive compounds, and increased superoxide dismutase (SOD), catalase (CAT), and peroxidase (POD) activity.

  5. Optimization of extraction of β-endoglucanase from the fermented bran of Aspergillus niger.

    Science.gov (United States)

    Chandra, M Subhosh; Viswanath, Buddolla; Reddy, B Rajasekhar

    2010-10-01

    A local isolate of Aspergillus niger was cultivated under optimal growth conditions on wheat bran in solid state fermentation. β-endoglucanase from fermented bran was separately extracted with different solvents to test recovery of enzyme. Among solvents tested, distilled water served the best leachate. Conditions were further optimized with this leachate. Two washes of fermented bran with the leachate for 30 min each under shaking conditions in a ratio of 1 g of wheat bran: 4 ml of distilled water together yielded maximum recovery of 16.7 U/g of wheat bran.

  6. Xylanase production by a local fungal isolate, Aspergillus niger USM AI 1 via solid state

    OpenAIRE

    Ibrahim Che Omar; Pang Pei Kheng

    2005-01-01

    Isolate USM A1 I which was identified to be Aspergillus niger was selected as a potential producer of xylanase via a solid state fermentation system (SSF) using palm kernel cake (PKC) as substrate. The modification of the physical conditions of the SSF system indicated that the xylanase activity was 23.97 U/g PKC at the moisture ratio of 1:0.75 of PKC: moistening agent with the inoculum size of 1¥104 spores/ml and cultivated at the ambient temperature (28±3ºC). The supplementation of addition...

  7. Dephosphorylation of Phytate by Using the Aspergillus niger Phytase with a High Affinity for Phytate

    OpenAIRE

    Nagashima, Tadashi; Tange, Tatsuya; Anazawa, Hideharu

    1999-01-01

    A phytase (EC 3.1.3.8) with a high affinity for phytic acid was found in Aspergillus niger SK-57 and purified to homogeneity in four steps by using ion-exchange chromatography (two types), gel filtration, and chromatofocusing. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of the purified enzyme gave a single stained band at a molecular mass of approximately 60 kDa. The Michaelis constant of the enzyme for phytic acid (18.7 ± 4.6 μM) was statistically analyzed. In regard to the ort...

  8. Optimization of solid state fermentation of sugar cane by Aspergillus niger considering particles size effect

    Energy Technology Data Exchange (ETDEWEB)

    Echevarria, J.; Rodriguez, L.J.A.; Delgado, G. (Instituto Cubano de Investigaciones de los Derivados de la Cana de Azucar (ICIDCA), La Habana (Cuba)); Espinosa, M.E. (Centro Nacional de Investigaciones Cientificas, La Habana (Cuba))

    1991-01-01

    The protein enrichment of sugar cane by solid state fermentation employing Aspergillus niger was optimized in a packed bed column using a two Factor Central Composit Design {alpha} = 2, considering as independent factors the particle diameter corresponding to different times of grinding for a sample and the air flow rate. It was significative for the air flow rate (optimum 4.34 VKgM) and the particle diameter (optimum 0.136 cm). The average particle size distribution, shape factor, specific surface, volume-surface mean diameter, number of particles, real and apparent density and holloweness for the different times of grinding were determined, in order to characterize the samples. (orig.).

  9. Indole-3-acetic acid and gibberellic acid production in Aspergillus niger

    OpenAIRE

    BİLKAY, Işıl SEYİS; KARAKOÇ, Şafak; AKSÖZ, Nilüfer

    2010-01-01

    The effects of incubation time, temperature, pH, and agitation on indole-3- acetic acid and gibberellic acid production in Aspergillus niger were studied. For indole-3-acetic acid production, 6 days of incubation at 25 °C and pH 6.0 was found to be optimum. Optimum conditions for gibberellic acid production were 12 days of incubation at 30 °C and pH 5.0. Agitation increased both indole-3-acetic acid and gibberellic acid production.

  10. Production of Citric Acid from a New Substrate, Undersized Semolina, by Aspergillus niger

    OpenAIRE

    Emine Alben; Osman Erkmen

    2004-01-01

    The production of citric acid from fermentation medium (mass per volume ratio 0.01 % of undersized semolina) by Aspergillus niger was studied by shake culture method. The effects of initial pH (4.5, 5.5 and 6.5), methanol (volume fraction 1.0, 2.0 and 3.0 %) and ammonium nitrate (mass per volume ratio 0.01 %) on the production of citric acid were investigated. Citric acid concentration, biomass concentration and the amount of total carbohydrates (as glucose) were determined during fermentatio...

  11. Rewiring a secondary metabolite pathway towards itaconic acid production in Aspergillus niger.

    Science.gov (United States)

    Hossain, Abeer H; Li, An; Brickwedde, Anja; Wilms, Lars; Caspers, Martien; Overkamp, Karin; Punt, Peter J

    2016-07-28

    The industrially relevant filamentous fungus Aspergillus niger is widely used in industry for its secretion capabilities of enzymes and organic acids. Biotechnologically produced organic acids promise to be an attractive alternative for the chemical industry to replace petrochemicals. Itaconic acid (IA) has been identified as one of the top twelve building block chemicals which have high potential to be produced by biotechnological means. The IA biosynthesis cluster (cadA, mttA and mfsA) has been elucidated in its natural producer Aspergillus terreus and transferred to A. niger to enable IA production. Here we report the rewiring of a secondary metabolite pathway towards further improved IA production through the overexpression of a putative cytosolic citrate synthase citB in a A. niger strain carrying the IA biosynthesis cluster. We have previously shown that expression of cadA from A. terreus results in itaconic acid production in A. niger AB1.13, albeit at low levels. This low-level production is boosted fivefold by the overexpression of mttA and mfsA in itaconic acid producing AB1.13 CAD background strains. Controlled batch cultivations with AB1.13 CAD + MFS + MTT strains showed increased production of itaconic acid compared with AB1.13 CAD strain. Moreover, preliminary RNA-Seq analysis of an itaconic acid producing AB1.13 CAD strain has led to the identification of the putative cytosolic citrate synthase citB which was induced in an IA producing strain. We have overexpressed citB in a AB1.13 CAD + MFS + MTT strain and by doing so hypothesize to have targeted itaconic acid production to the cytosolic compartment. By overexpressing citB in AB1.13 CAD + MFS + MTT strains in controlled batch cultivations we have achieved highly increased titers of up to 26.2 g/L IA with a productivity of 0.35 g/L/h while no CA was produced. Expression of the IA biosynthesis cluster in Aspergillus niger AB1.13 strain enables IA production. Moreover, in the AB1.13 CAD

  12. Aspergillus niger membrane-associated proteome analysis for the identification of glucose transporters.

    Science.gov (United States)

    Sloothaak, J; Odoni, D I; de Graaff, L H; Martins Dos Santos, V A P; Schaap, P J; Tamayo-Ramos, J A

    2015-01-01

    The development of biological processes that replace the existing petrochemical-based industry is one of the biggest challenges in biotechnology. Aspergillus niger is one of the main industrial producers of lignocellulolytic enzymes, which are used in the conversion of lignocellulosic feedstocks into fermentable sugars. Both the hydrolytic enzymes responsible for lignocellulose depolymerisation and the molecular mechanisms controlling their expression have been well described, but little is known about the transport systems for sugar uptake in A. niger. Understanding the transportome of A. niger is essential to achieve further improvements at strain and process design level. Therefore, this study aims to identify and classify A. niger sugar transporters, using newly developed tools for in silico and in vivo analysis of its membrane-associated proteome. In the present research work, a hidden Markov model (HMM), that shows a good performance in the identification and segmentation of functionally validated glucose transporters, was constructed. The model (HMMgluT) was used to analyse the A. niger membrane-associated proteome response to high and low glucose concentrations at a low pH. By combining the abundance patterns of the proteins found in the A. niger plasmalemma proteome with their HMMgluT scores, two new putative high-affinity glucose transporters, denoted MstG and MstH, were identified. MstG and MstH were functionally validated and biochemically characterised by heterologous expression in a S. cerevisiae glucose transport null mutant. They were shown to be a high-affinity glucose transporter (K m = 0.5 ± 0.04 mM) and a very high-affinity glucose transporter (K m = 0.06 ± 0.005 mM), respectively. This study, focusing for the first time on the membrane-associated proteome of the industrially relevant organism A. niger, shows the global response of the transportome to the availability of different glucose concentrations. Analysis of the A. niger

  13. PENAMBAHAN MIKROBA, Aspergillus niger DALAM BUNGKIL KELAPA SAWIT SEBAGAI BAHAN BAKU PAKAN UNTUK PEMBESARAN IKAN KERAPU MACAN

    Directory of Open Access Journals (Sweden)

    Neltje Nobertine Palinggi

    2008-12-01

    Full Text Available Penelitian ini dilakukan untuk mendapatkan informasi tentang pengaruh dosis Aspergillus niger dalam bungkil kelapa sawit sebagai bahan pakan pada pembesaran ikan kerapu macan. Ikan uji yang digunakan berukuran bobot rata-rata 23,15±0,23 g; ditebar dalam keramba jaring apung ukuran 1 m x 1 m x 2 m, dengan kepadatan 16 ekor/keramba. Perlakuan yang diuji adalah penambahan Aspergillus niger sebanyak 2, 4, 8, 16 g/kg bungkil kelapa sawit dan kontrol. Masing-masing perlakuan diulang tiga kali dan disain adalah rancangan acak lengkap. Selama pemeliharaan, ikan diberi pakan uji dua kali sehari (pagi dan sore secara satiasi selama 20 minggu. Hasil penelitian menunjukkan bahwa penambahan 8 g Aspergillus niger/kg bungkil kelapa sawit memberikan pertambahan bobot dan laju spesifik lebih tinggi daripada kontrol (P0,05, namun nilainya nyata lebih tinggi (P0.05 among those of the juveniles fed on the diets with 2, 4, 16 g of A. niger/kg palm oil cake. Although the feed efficiency, protein efficiency ratio and protein retention of juveniles fed on the diet with 8 g A. niger/kg palm oil cake were not significantly different (P>0.05 from those of the juveniles fed on the diets with 2 and 4 g of A. niger/kg palm oil cake, those of juveniles the fed diet with 8 g of A. niger/kg palm oil cake were significantly higher (P<0.05 than those of the juveniles fed the diet with 16 g A. niger/kg palm oil cake. The best of growth rate of tiger grouper juveniles occurred at the dosage of 7.8—8.2 g A. niger/kg palm oil cake.

  14. The cell wall of the filamentous fungus Aspergillus niger

    NARCIS (Netherlands)

    Damveld, Robbert A.

    2005-01-01

    Fungi are a very successful species and are distributed worldwide. However, the presence of fungi is not always desired. Filamentous fungi can grow on living or dead organic material and even inside the host. Current methods to prevent fungal growth are insufficient, causing fatality after fungal

  15. Protein secretion in the filamentous fungus Aspergillus niger

    NARCIS (Netherlands)

    Weenink, Xavier Oswin

    2008-01-01

    Filamentous fungi are multicellular eukaryotic organisms, which represent a separate taxonomic group organisms within the fungal kingdom, apart from the yeasts. These fungi always need a substrate to grow on, this can be living or dead material. Fungi possess the capacity to secrete high levels of

  16. Functional genomics analysis of the secretory pathway in Aspergillus niger

    NARCIS (Netherlands)

    Oliveira, J.M.

    2010-01-01

    Filamentous fungi can be found in the majority of habitats of our planet. The wide-spread presence of filamentous fungi is related to their versatile metabolism, which allows them to grow on simple substrates, such as nitrate, acetate, ethanol, ammonia, or on complex matter such as biopolymers from

  17. Comparative genomics and transcriptome analysis of Aspergillus niger and metabolic engineering for citrate production

    Science.gov (United States)

    Yin, Xian; Shin, Hyun-dong; Li, Jianghua; Du, Guocheng; Liu, Long; Chen, Jian

    2017-01-01

    Despite a long and successful history of citrate production in Aspergillus niger, the molecular mechanism of citrate accumulation is only partially understood. In this study, we used comparative genomics and transcriptome analysis of citrate-producing strains—namely, A. niger H915-1 (citrate titer: 157 g L−1), A1 (117 g L−1), and L2 (76 g L−1)—to gain a genome-wide view of the mechanism of citrate accumulation. Compared with A. niger A1 and L2, A. niger H915-1 contained 92 mutated genes, including a succinate-semialdehyde dehydrogenase in the γ-aminobutyric acid shunt pathway and an aconitase family protein involved in citrate synthesis. Furthermore, transcriptome analysis of A. niger H915-1 revealed that the transcription levels of 479 genes changed between the cell growth stage (6 h) and the citrate synthesis stage (12 h, 24 h, 36 h, and 48 h). In the glycolysis pathway, triosephosphate isomerase was up-regulated, whereas pyruvate kinase was down-regulated. Two cytosol ATP-citrate lyases, which take part in the cycle of citrate synthesis, were up-regulated, and may coordinate with the alternative oxidases in the alternative respiratory pathway for energy balance. Finally, deletion of the oxaloacetate acetylhydrolase gene in H915-1 eliminated oxalate formation but neither influence on pH decrease nor difference in citrate production were observed. PMID:28106122

  18. Genome mining and functional genomics for siderophore production in Aspergillus niger.

    Science.gov (United States)

    Franken, Angelique C W; Lechner, Beatrix E; Werner, Ernst R; Haas, Hubertus; Lokman, B Christien; Ram, Arthur F J; van den Hondel, Cees A M J J; de Weert, Sandra; Punt, Peter J

    2014-11-01

    Iron is an essential metal for many organisms, but the biologically relevant form of iron is scarce because of rapid oxidation resulting in low solubility. Simultaneously, excessive accumulation of iron is toxic. Consequently, iron uptake is a highly controlled process. In most fungal species, siderophores play a central role in iron handling. Siderophores are small iron-specific chelators that can be secreted to scavenge environmental iron or bind intracellular iron with high affinity. A second high-affinity iron uptake mechanism is reductive iron assimilation (RIA). As shown in Aspergillus fumigatus and Aspergillus nidulans, synthesis of siderophores in Aspergilli is predominantly under control of the transcription factors SreA and HapX, which are connected by a negative transcriptional feedback loop. Abolishing this fine-tuned regulation corroborates iron homeostasis, including heme biosynthesis, which could be biotechnologically of interest, e.g. the heterologous production of heme-dependent peroxidases. Aspergillus niger genome inspection identified orthologues of several genes relevant for RIA and siderophore metabolism, as well as sreA and hapX. Interestingly, genes related to synthesis of the common fungal extracellular siderophore triacetylfusarinine C were absent. Reverse-phase high-performance liquid chromatography (HPLC) confirmed the absence of triacetylfusarinine C, and demonstrated that the major secreted siderophores of A. niger are coprogen B and ferrichrome, which is also the dominant intracellular siderophore. In A. niger wild type grown under iron-replete conditions, the expression of genes involved in coprogen biosynthesis and RIA was low in the exponential growth phase but significantly induced during ascospore germination. Deletion of sreA in A. niger resulted in elevated iron uptake and increased cellular ferrichrome accumulation. Increased sensitivity toward phleomycin and high iron concentration reflected the toxic effects of excessive

  19. Optimization of catalase biosynthesis in submerged cultures of Aspergillus niger mutant.

    Science.gov (United States)

    Gromada, A; Fiedurek, J

    1997-01-01

    The effect of some medium components, viscous substances and metabolic inhibitors, on catalase production by mutant Aspergillus niger has been studied in shake culture. Altering the composition of the basal medium, particularly substituting NaNO3 for KNO3, and peptone for yeast extract brought an increase in extra- and intracellular catalase activity by 1.5- and 3-fold, respectively. The addition of 2.0-6.0 mg sodium alginate or pectin/ml as viscous additive to the medium, containing glucose as carbon source, increased the medium viscosity and catalase production in shake culture by about 2.8- to 3.0-fold. The highest yield of extracellular catalase activity of A. niger was obtained in the presence of sodium orthovanadate and Triton X-100, which improved the activity of this enzyme by about 1.5-2.2-fold. A significant increase in intracellular catalase activity was observed in the presence of hematin, Tween 80 and sodium orthovanadate (1.7-, 1.6- and 1.4-fold respectively). The time course of growth and enzyme production by A. niger in the optimized medium is also reported.

  20. Efficacy and possible mechanisms of perillaldehyde in control of Aspergillus niger causing grape decay.

    Science.gov (United States)

    Tian, Jun; Wang, Yanzhen; Zeng, Hong; Li, Zongyun; Zhang, Peng; Tessema, Akalate; Peng, Xue

    2015-06-02

    A variety of plant products have been recognized for their antifungal activity and recently have attracted food industry attention for their efficacy in controlling postharvest fungal decay of fruits. The antifungal activity of perillaldehyde (PAE) was evaluated against Aspergillus niger, a known cause of grape spoilage, and possible mechanisms were explored. PAE showed notable antifungal activity against A. niger, with a minimum inhibitory concentration (MIC) and a minimum fungicidal concentration (MFC) of 0.25 and 1 μl/ml, respectively. The accumulation of mycelial biomass was also inhibited by PAE in a dose-dependent manner, completely inhibiting mycelial growth at 1 μl/ml. In vivo data confirmed that the vapour treatment of grapes with various concentrations of PAE markedly improved control of A. niger and suppressed natural decay. Concentrations of PAE of 0.075 μl/ml air showed the greatest inhibition of fungal growth compared to the controls. Further experiments indicated that PAE activated a membrane-active mechanism that inhibits ergosterol synthesis, increases membrane permeability (as evidenced by extracellular pH and conductivity measurements), and disrupts membrane integrity, leading to cell death. Our findings suggest that this membrane-active mechanism makes PAE a promising potential antifungal agent for postharvest control of grape spoilage. Copyright © 2015 Elsevier B.V. All rights reserved.

  1. Regulation of the alpha-glucuronidase-encoding gene ( aguA) from Aspergillus niger.

    Science.gov (United States)

    de Vries, R P; van de Vondervoort, P J I; Hendriks, L; van de Belt, M; Visser, J

    2002-09-01

    The alpha-glucuronidase gene aguA from Aspergillus niger was cloned and characterised. Analysis of the promoter region of aguA revealed the presence of four putative binding sites for the major carbon catabolite repressor protein CREA and one putative binding site for the transcriptional activator XLNR. In addition, a sequence motif was detected which differed only in the last nucleotide from the XLNR consensus site. A construct in which part of the aguA coding region was deleted still resulted in production of a stable mRNA upon transformation of A. niger. The putative XLNR binding sites and two of the putative CREA binding sites were mutated individually in this construct and the effects on expression were examined in A. niger transformants. Northern analysis of the transformants revealed that the consensus XLNR site is not actually functional in the aguA promoter, whereas the sequence that diverges from the consensus at a single position is functional. This indicates that XLNR is also able to bind to the sequence GGCTAG, and the XLNR binding site consensus should therefore be changed to GGCTAR. Both CREA sites are functional, indicating that CREA has a strong influence on aguA expression. A detailed expression analysis of aguA in four genetic backgrounds revealed a second regulatory system involved in activation of aguA gene expression. This system responds to the presence of glucuronic and galacturonic acids, and is not dependent on XLNR.

  2. Novel Antifungal Peptides Produced by Leuconostoc mesenteroides DU15 Effectively Inhibit Growth of Aspergillus niger.

    Science.gov (United States)

    Muhialdin, Belal J; Hassan, Zaiton; Abu Bakar, Fatimah; Algboory, Hussein L; Saari, Nazamid

    2015-05-01

    The ability of Leuconostoc mesenteroides DU15 to produce antifungal peptides that inhibit growth of Aspergillus niger was evaluated under optimum growth conditions of 30 °C for 48 h. The cell-free supernatant showed inhibitory activity against A. niger. Five novel peptides were isolated with the sequences GPFPL, YVPLF, LLHGVPLP, GPFPLEMTLGPT, and TVYPFPGPL as identified by de novo sequencing using PEAKS 6 software. Peptide LLHGVPLP was the only positively charged (cationic peptides) and peptide GPFPLEMTLGPT negatively charged (anionic), whereas the rest are neutral. The identified peptides had high hydrophobicity ratio and low molecular weights with amino acids sequences ranging from 5 to 12 residues. The mode of action of these peptides is observed under the scanning electron microscope and is due to cell lysis of fungi. This work reveals the potential of peptides from L. mesenteroides DU15 as natural antifungal preservatives in inhibiting the growth of A. niger that is implicated to the spoilage during storage. © 2015 Institute of Food Technologists®

  3. Antifungal mechanism of antibacterial peptide, ABP-CM4, from Bombyx mori against Aspergillus niger.

    Science.gov (United States)

    Zhang, Jie; Wu, Xi; Zhang, Shuang-Quan

    2008-12-01

    Antibacterial peptide, CM4 (ABP-CM4), a 35 amino acid peptide from Chinese silkworm-Bombyx mori, displayed a strong antifungal activity against Aspergillus niger, Trichoderma viride and Gibberella saubinetii. Scanning electron microcopy showed that the morphology of conidia became more irregular and swelled when treated with ABP-CM4 at its minimal inhibitory concentration (MIC) of 8 muM. A cell wall regeneration assay indicated that the plasma membrane was the prime target of ABP-CM4 action. Confocal laser scanning microscopy showed that the cytoskeleton of A. niger was destroyed when treated with ABP-CM4 at 8 muM. Furthermore, transmission electron microscopy showed that the membrane and the cellular organelles of fungus were disrupted and there were many vacuoles in the fungal cellular space after the treatment with ABP-CM4. A gel-retardation assay showed that ABP-CM4 bound the DNA of A. niger. Our results suggest that ABP-CM4 exerts its antifungal activity by disrupting the structure of cell membranes and the cytoskeleton and interacts with the organelles, such as the mitochondrion and with the DNA in the fungal cell, subsequently resulting in cell death.

  4. Putative Aspergillus niger-induced oxalate nephrosis in sheep : clinical communication

    Directory of Open Access Journals (Sweden)

    C.J. Botha

    2009-05-01

    Full Text Available A sheep farmer provided a maize-based brewer's grain (mieliemaroek and bales of Eragrostis curvula hay to ewes and their lambs, kept on zero-grazing in pens. The 'mieliemaroek' was visibly mouldy. After 14 days in the feedlot, clinical signs, including generalised weakness, ataxia of the hind limbs, tremors and recumbency, were noticed. Six ewes died within a period of 7 days. A post mortem examination was performed on 1 ewe. The carcass appeared to be cachectic with mild effusions into the body cavities; mild lung congestion and pallor of the kidneys were observed. Microscopical evaluation revealed nephrosis and birefringent oxalate crystals in the renal tubules when viewed under polarised light. A provisional diagnosis of oxalate nephrosis with subsequent kidney failure was made. Amongst other fungi, Aspergillus niger was isolated from 'mieliemaroek' samples submitted for fungal culture and identification. As A. niger is known to synthesise oxalates, a qualitative screen to detect oxalic acid in the mieliemaroek and purified A. niger isolates was performed using high-performance liquid chromatography (HPLC. Oxalic acid was detected, which supported a diagnosis of soluble oxalate-induced nephropathy.

  5. Removal and recovery of uranium (VI) from aqueous solutions by immobilized Aspergillus niger powder beads.

    Science.gov (United States)

    Ding, De-Xin; Tan, Xiang; Hu, Nan; Li, Guang-Yue; Wang, Yong-Dong; Tan, Yan

    2012-11-01

    The immobilized Aspergillus niger powder beads were obtained by entrapping nonviable A. niger powder into Ca-alginate gel. The effects of pH, contact time, initial uranium (VI) concentration and biomass dosage on the biosorption of uranium (VI) onto the beads from aqueous solutions were investigated in a batch system. Biosorption equilibrium data were agreeable with Langmuir isotherm model and the maximum biosorption capacity of the beads for uranium (VI) was estimated to be 649.4 mg/g at 30 °C. The biosorption kinetics followed the pseudo-second-order model and intraparticle diffusion equation. The variations in enthalpy (26.45 kJ/mol), entropy (0.167 kJ/mol K) and Gibbs free energy were calculated from the experimental data. SEM and EDS analysis indicated that the beads have strong adsorption capability for uranium (VI). The adsorbed uranium (VI) on the beads could be released with HNO(3) or HCl. The results showed that the immobilized A. niger powder beads had great potential for removing and recovering uranium (VI) from aqueous solutions.

  6. Physicochemical Properties Analysis and Secretome of Aspergillus niger in Fermented Rapeseed Meal.

    Directory of Open Access Journals (Sweden)

    Changyou Shi

    Full Text Available The nutrient digestibility and feeding value of rapeseed meal (RSM for non-ruminant animals is poor due to the presence of anti-nutritional substances such as glucosinolate, phytic acid, crude fiber etc. In the present study, a solid state fermentation (SSF using Aspergillus niger was carried out with the purpose of improving the nutritional quality of RSM. The chemical composition and physicochemical properties of RSM before and after fermentation were compared. To further understand possible mechanism of solid state fermentation, the composition of extracellular enzymes secreted by Aspergillus niger during fermentation was analysed using two-dimentional difference gel electrophoresis (2D-DIGE combined with matrix assisted laser desorption ionization-time of flight-mass spectrometer (MALDI-TOF-MS. Results of the present study indicated that SSF had significant effects on chemical composition of RSM. The fermented rapeseed meal (FRSM contained more crude protein (CP and amino acid (AA (except His than unfermented RSM. Notably, the small peptide in FRSM was 2.26 time larger than that in unfermented RSM. Concentrations of anti-nutritional substrates in FRSM including neutral detergent fiber (NDF, glucosinolates, isothiocyanate, oxazolidithione, and phytic acid declined (P < 0.05 by 13.47, 43.07, 55.64, 44.68 and 86.09%, respectively, compared with unfermented RSM. A. niger fermentation disrupted the surface structure, changed macromolecular organic compounds, and reduced the protein molecular weights of RSM substrate. Total proteins of raw RSM and FRSM were separated and 51 protein spots were selected for mass spectrometry according to 2D-DIGE map. In identified proteins, there were 15 extracellular hydrolases secreted by A. niger including glucoamylase, acid protease, beta-glucanase, arabinofuranosidase, xylanase, and phytase. Some antioxidant related enzymes also were identified. These findings suggested that A. niger is able to secrete many

  7. Chemical mimicking of bio-assisted aluminium extraction by Aspergillus niger's exometabolites.

    Science.gov (United States)

    Boriová, Katarína; Urík, Martin; Bujdoš, Marek; Pifková, Ivana; Matúš, Peter

    2016-11-01

    Presence of microorganisms in soils strongly affects mobility of metals. This fact is often excluded when mobile metal fraction in soil is studied using extraction procedures. Thus, the first objective of this paper was to evaluate strain Aspergillus niger's exometabolites contribution on aluminium mobilization. Fungal exudates collected in various time intervals during cultivation were analyzed and used for two-step bio-assisted extraction of alumina and gibbsite. Oxalic, citric and gluconic acids were identified in collected culture media with concentrations up to 68.4, 2.0 and 16.5 mmol L-1, respectively. These exometabolites proved to be the most efficient agents in mobile aluminium fraction extraction with aluminium extraction efficiency reaching almost 2.2%. However, fungal cultivation is time demanding process. Therefore, the second objective was to simplify acquisition of equally efficient extracting agent by chemically mimicking composition of main organic acid components of fungal exudates. This was successfully achieved with organic acids mixture prepared according to medium composition collected on the 12th day of Aspergillus niger cultivation. This mixture extracted similar amounts of aluminium from alumina compared to culture medium. The aluminium extraction efficiency from gibbsite by organic acids mixture was lesser than 0.09% which is most likely because of more rigid mineral structure of gibbsite compared to alumina. The prepared organic acid mixture was then successfully applied for aluminium extraction from soil samples and compared to standard single step extraction techniques. This showed there is at least 2.9 times higher content of mobile aluminium fraction in soils than it was previously considered, if contribution of microbial metabolites is considered in extraction procedures. Thus, our contribution highlights the significance of fungal metabolites in aluminium extraction from environmental samples, but it also simplifies the extraction

  8. Enhanced itaconic acid production in Aspergillus niger using genetic modification and medium optimization

    Directory of Open Access Journals (Sweden)

    Li An

    2012-08-01

    Full Text Available Abstract Background Aspergillus niger was selected as a host for producing itaconic acid due to its versatile and tolerant character in various growth environments, and its extremely high capacity of accumulating the precursor of itaconic acid: citric acid. Expressing the CAD gene from Aspergillus terreus opened the metabolic pathway towards itaconic acid in A. niger. In order to increase the production level, we continued by modifying its genome and optimizing cultivation media. Results Based on the results of previous transcriptomics studies and research from other groups, two genes : gpdA encoding the glyceraldehyde −3-dehydrogenase (GPD and hbd1 encoding a flavohemoglobin domain (HBD were overexpressed in A. niger. Besides, new media were designed based on a reference medium for A. terreus. To analyze large numbers of cultures, we developed an approach for screening both fungal transformants and various media in 96-well micro-titer plates. The hbd1 transformants (HBD 2.2/2.5 did not improve itaconic acid titer while the gpdA transformant (GPD 4.3 decreased the itaconic acid production. Using 20 different media, copper was discovered to have a positive influence on itaconic acid production. Effects observed in the micro-titer plate screening were confirmed in controlled batch fermentation. Conclusions The performance of gpdA and hbd1 transformants was found not to be beneficial for itaconic acid production using the tested cultivation conditions. Medium optimization showed that, copper was positively correlated with improved itaconic acid production. Interestingly, the optimal conditions for itaconic acid clearly differ from conditions optimal for citric- and oxalic acid production.

  9. Enhanced itaconic acid production in Aspergillus niger using genetic modification and medium optimization.

    Science.gov (United States)

    Li, An; Pfelzer, Nina; Zuijderwijk, Robbert; Punt, Peter

    2012-08-27

    Aspergillus niger was selected as a host for producing itaconic acid due to its versatile and tolerant character in various growth environments, and its extremely high capacity of accumulating the precursor of itaconic acid: citric acid. Expressing the CAD gene from Aspergillus terreus opened the metabolic pathway towards itaconic acid in A. niger. In order to increase the production level, we continued by modifying its genome and optimizing cultivation media. Based on the results of previous transcriptomics studies and research from other groups, two genes : gpdA encoding the glyceraldehyde -3-dehydrogenase (GPD) and hbd1 encoding a flavohemoglobin domain (HBD) were overexpressed in A. niger. Besides, new media were designed based on a reference medium for A. terreus. To analyze large numbers of cultures, we developed an approach for screening both fungal transformants and various media in 96-well micro-titer plates. The hbd1 transformants (HBD 2.2/2.5) did not improve itaconic acid titer while the gpdA transformant (GPD 4.3) decreased the itaconic acid production. Using 20 different media, copper was discovered to have a positive influence on itaconic acid production. Effects observed in the micro-titer plate screening were confirmed in controlled batch fermentation. The performance of gpdA and hbd1 transformants was found not to be beneficial for itaconic acid production using the tested cultivation conditions. Medium optimization showed that, copper was positively correlated with improved itaconic acid production. Interestingly, the optimal conditions for itaconic acid clearly differ from conditions optimal for citric- and oxalic acid production.

  10. Production of biologically active oxidized derivatives of finasteride through metabolism by Aspergillus niger culture.

    Science.gov (United States)

    Ali, Sajid; Nisar, Muhammad; Shah, Zarbad

    2016-11-01

    Among the 4-azasteroids, finasteride is biologically the most important compound having preventive effect against male pattern baldness (MPH) and benign prostatic hyperplasia commonly called enlargement of prostate gland. The microbial transformation of finasteride by fungus Aspergillus niger (ATCC 10549) has been investigated to obtain biologically more potent derivatives. Fermentation of finasteride was performed with filamentous fungus Aspergillus niger (ATCC 10549). This transformation resulted in the production of two transformed products, which were purified through column chromatography. In vitro lipoxygenase inhibitory potential was determined by incubating 20 mL of the enzyme with 10 mL of test sample (100 μM) in 0.1 mM (pH 7.0) phosphate buffer for 5 min at 258 °C followed by addition of 10 μL of substrate (linolenic acid) to reaction mixture and measuring the formation of complex spectrophotometrically. Structure elucidation of biotransformed metabolites was ascertained through extensive 1D and 2D spectroscopic techniques. This study established the fact that A. niger promoted stereospecific dihydroxylation at C-11 and C-15 of finasteride. The resulting biotransformed metabolites were characterized as 11α-hydroxyfinasteride and 15β-hydroxyfinasteride, respectively. Finasteride along with transformed metabolites were analyzed for their in vitro lipoxygenase (LOX) inhibition assay. Among the tested compounds 15β-hydroxyfinasteride showed good activity with IC50 value 112.56 ± 2.23 μM while inhibitory effect in case of 11α-hydroxyfinasteride was low with IC50 value 186.05 ± 1.34 μM. Standard compound baicalein revealed IC50 value being 22.0 ± 0.05 μM. The present investigation highlighted the fact that potentially active compound can be produced through the technology of biotransformation.

  11. Enhanced itaconic acid production in Aspergillus niger using genetic modification and medium optimization

    Science.gov (United States)

    2012-01-01

    Background Aspergillus niger was selected as a host for producing itaconic acid due to its versatile and tolerant character in various growth environments, and its extremely high capacity of accumulating the precursor of itaconic acid: citric acid. Expressing the CAD gene from Aspergillus terreus opened the metabolic pathway towards itaconic acid in A. niger. In order to increase the production level, we continued by modifying its genome and optimizing cultivation media. Results Based on the results of previous transcriptomics studies and research from other groups, two genes : gpdA encoding the glyceraldehyde −3-dehydrogenase (GPD) and hbd1 encoding a flavohemoglobin domain (HBD) were overexpressed in A. niger. Besides, new media were designed based on a reference medium for A. terreus. To analyze large numbers of cultures, we developed an approach for screening both fungal transformants and various media in 96-well micro-titer plates. The hbd1 transformants (HBD 2.2/2.5) did not improve itaconic acid titer while the gpdA transformant (GPD 4.3) decreased the itaconic acid production. Using 20 different media, copper was discovered to have a positive influence on itaconic acid production. Effects observed in the micro-titer plate screening were confirmed in controlled batch fermentation. Conclusions The performance of gpdA and hbd1 transformants was found not to be beneficial for itaconic acid production using the tested cultivation conditions. Medium optimization showed that, copper was positively correlated with improved itaconic acid production. Interestingly, the optimal conditions for itaconic acid clearly differ from conditions optimal for citric- and oxalic acid production. PMID:22925689

  12. Deletion of a Chitin Synthase Gene in a Citric Acid Producing Strain of Aspergillus niger

    Energy Technology Data Exchange (ETDEWEB)

    Rinker, Torri E.; Baker, Scott E.

    2007-01-29

    Citric acid production by the filamentous fungus Aspergillus niger is carried out in a process that causes the organism to drastically alter its morphology. This altered morphology includes hyphal swelling and highly limited polar growth resulting in clumps of swollen cells that eventually aggregate into pellets of approximately 100 microns in diameter. In this pelleted form, A. niger has increased citric acid production as compared to growth in filamentous form. Chitin is a crucial component of the cell wall of filamentous fungi. Alterations in the deposition or production of chitin may have profound effects on the morphology of the organism. In order to study the role of chitin synthesis in pellet formation we have deleted a chitin synthase gene (csmA) in Aspergillus niger strain ATCC 11414 using a PCR based deletion construct. This class of chitin synthases is only found in filamentous fungi and is not present in yeasts. The csmA genes contain a myosin motor domain at the N-terminus and a chitin synthesis domain at the C-terminus. They are believed to contribute to the specialized polar growth observed in filamentous fungi that is lacking in yeasts. The csmA deletion strain (csmAΔ) was subjected to minimal media with and without osmotic stabilizers as well as tested in citric acid production media. Without osmotic stabilizers, the mutant germlings were abnormally swollen, primarily in the subapical regions, and contained large vacuoles. However, this swelling is ultimately not inhibitory to growth as the germlings are able to recover and undergo polar growth. Colony formation was largely unaffected in the absence of osmotic stabilizers. In citric acid production media csmAΔ was observed to have a 2.5 fold increase in citric acid production. The controlled expression of this class of chitin synthases may be useful for improving production of organic acids in filamentous fungi.

  13. Tratamento de efluentes de refinaria de petróleo em reatores com Aspergillus niger Treatment of petroleum refinery wastewater by reactors inoculated with Aspergillus niger

    Directory of Open Access Journals (Sweden)

    Sandra Tédde Santaella

    2009-03-01

    Full Text Available Neste trabalho, avaliou-se o efeito do tempo de detenção hidráulica (TDH no desempenho de três reatores aeróbios inoculados com Aspergillus niger AN400, usados para tratamento de efluentes de refinarias de petróleo. Cada reator foi operado com um tempo de detenção hidráulica diferente: 4, 8 e 12 horas, durante 152 dias. Eles possuíam leito fixo de espuma de poliuretano e o escoamento era ascendente e contínuo. Determinaram-se: pH, fenóis, demanda química de oxigênio (DQO, amônia, nitrito e nitrato, no afluente e efluentes dos reatores. O TDH de oito horas foi o melhor para remoção de DQOsolúvel e não houve diferença entre os TDHs para remoção de fenóis totais. No período estável não houve remoção de nitrato; no entanto ocorreu remoção de nitrito de aproximadamente 99%. Além disto, houve produção de amônia devido à amonificação a partir do nitrito presente no meio.This paper evaluated the effect of hydraulic retention time (HRT on the performance of three upflow aerobic reactors, with polyurethane foam as support material, inoculated with Aspergillus niger AN400, used for the treatment of petroleum refinery wastewater. Each reactor was operated with a different HRT: 4, 8 and 12 hours, during 152 days. The performance was evaluated based on pH; phenols; COD, nitrate and nitrite. The results show that for the COD removal, it is more reasonable to operate the reactor with HRT of eight hours. However, there was no difference among results of phenol removal efficiency of the different HRTs. During steady state condition, nitrite was removed in approximately 99%, but there was no reduction on the nitrate concentration. Ammonia was produced in all reactors, probably due to ammonification of nitrite.

  14. The FlbA-regulated predicted transcription factor Fum21 of Aspergillus niger is involved in fumonisin production

    DEFF Research Database (Denmark)

    Aerts, David; Hauer, Esther E.; Ohm, Robin A.

    2017-01-01

    Aspergillus niger secretes proteins throughout the colony except for the zone that forms asexual spores called conidia. Inactivation of flbA that encodes a regulator of G-protein signaling results in colonies that are unable to reproduce asexually and that secrete proteins throughout the mycelium...

  15. Decolorization and detoxification of Synozol red HF-6BN azo dye, by Aspergillus niger and Nigrospora sp

    Directory of Open Access Journals (Sweden)

    Sidra Ilyas

    2013-01-01

    Full Text Available In the present investigation the fungi, Aspergillus niger and Nigrospora sp. were employed for decolorization of Synazol red HF-6BN. Decolorization study showed that Aspergillus niger and Nigrospora sp. were able to decolorize 88% and 96% Synazol red 6BN, respectively, in 24 days. It was also studied that 86% and 90% Synazol red containing of dye effluent was decolorized by Aspergillus niger and Nigrospora sp. after 28 days of incubation at room temperature. A fungal-based protein with relative molecular mass of 70 kDa was partially purified and examined for enzymatic characteristics. The enzyme exhibited highest activity at temperature ranging from 40-50[degree sign]C and at pH=6.0. The enzyme activity was enhanced in the presence of metal cations. High performance liquid chromatography analysis confirmed that these fungal strains are capable to degrade Synazol red dye into metabolites. No zones of inhibition on agar plates and growth of Vigna radiata in the presence of dye extracted sample, indicated that the fungal degraded dye metabolites are nontoxic to beneficial micro-flora and plant growth. Aspergillus niger and Nigrospora sp. have promising potential in color removal from textile wastewater-containing azo dyes.

  16. Cloning and characterization of the NADPH cytochrome P450 oxidoreductase gene from the filamentous fungus Aspergillus niger

    NARCIS (Netherlands)

    Brink, J.M. van den; Zeijl, C.M.J. van; Brons, J.F.; Hondel, C.A.M.J.J. van den; Gorcom, R.F.M. van

    1995-01-01

    In this paper, we describe the cloning and molecular characterization of the Aspergillus niger cytochrome P450 reductase (CPR) gene, cprA. Attempts to clone the cprA gene by heterologous hybridization techniques were unsuccessful. Using the polymerase chain reaction (PCR) with degenerate primers

  17. PRODUCTION AND CHARACTERIZATION OF CELLULOLYTIC ENZYMES BY ASPERGILLUS NIGER AND RHIZOPUS SP . BY SOLID STATE FERMENTATION OF PRICKLY PEAR

    Directory of Open Access Journals (Sweden)

    TAMIRES CARVALHO DOS SANTOS

    2016-01-01

    Full Text Available Prickly palm cactus husk was used as a solid - state fermentation support substrate for the production of cellulolytic enzymes using Aspergillus niger and Rhizopus sp. A Box - Behnken design was used to evaluate the effects of water activity, fermentation time and temperature on endoglucanase and total cellulase production. Response Surface Methodology showed that optimum conditions for endoglucanase production were achieved at after 70.35 h of fermentation at 29.56°C and a water activity of 0.875 for Aspergillus niger and after 68.12 h at 30.41°C for Rhizopus sp. Optimum conditions for total cellulase production were achieved after 74.27 h of fermentation at 31.22°C for Aspergillus niger and after 72.48 h and 27.86°C for Rhizopus sp . Water activity had a significant effect on Aspergillus niger endoglucanase production only. In industrial applications, enzymatic characterization is important for optimizing variables such as temperature and pH. In this study we showed that endoglucanase and total cellulase had a high level of thermostability and pH stability in all the enzymatic extracts. Enzymatic deactivation kinetic experiments indicated that the enzymes remained active after the freezing of the crude extract. Based on the results, bioconversion of cactus is an excellent alternative for the production of thermostable enzymes.

  18. Radiosensitization of Aspergillus niger and Penicillium chrysogenum using basil essential oil and ionizing radiation for food decontamination.

    Science.gov (United States)

    The Minimum Inhibitory Concentration (MIC) of basil oil, was determined for two pathogenic fungi of rice, Aspergillus niger and Penicillium chrysogenum. The antifungal activity of the basil oil in combination with ionising radiation was then investigated to determine if basil oil caused radiosensit...

  19. Effect of temperature and water activity on the production of fumonisins by Aspergillus niger and different Fusarium species

    DEFF Research Database (Denmark)

    Mogensen, Jesper Mølgaard; Nielsen, Kristian Fog; Samson, Robert A.

    2009-01-01

    Background Fumonisins are economically important mycotoxins which until recently were considered to originate from only a few Fusarium species. However recently a putative fumonisin gene cluster was discovered in two different Aspergillus niger strains followed by detection of an actual fumonisin...

  20. Chitinases CtcB and CfcI modify the cell wall in sporulating aerial mycelium of Aspergillus niger

    NARCIS (Netherlands)

    van Munster, Jolanda M.; Nitsche, Benjamin M.; Krijgsheld, Pauline; van Wijk, Alle; Dijkhuizen, Lubbert; Wösten, Han A.; Ram, Arthur F.; van der Maarel, Marc J. E. C.

    Sporulation is an essential part of the life cycle of the industrially important filamentous fungus Aspergillus niger. The formation of conidiophores, spore-bearing structures, requires remodelling of the fungal cell wall, as demonstrated by the differences in carbohydrate composition of cell walls

  1. Effect of temperature and water activity on the production of fumonisins by Aspergillus niger and different Fusarium species

    NARCIS (Netherlands)

    Mogensen, J.M.; Nielsen, K.F.; Samson, R.A.; Frisvad, J.C.; Thrane, U.

    2009-01-01

    ABSTRACT: BACKGROUND: Fumonisins are economically important mycotoxins which until recently were considered to originate from only a few Fusarium species. However recently a putative fumonisin gene cluster was discovered in two different Aspergillus niger strains followed by detection of an actual

  2. EFFECT OF INOCULUM SIZE ON SURVIVAL RATE OF CANDIDA-ALBICANS AND ASPERGILLUS-NIGER IN TOPICAL PREPARATIONS

    NARCIS (Netherlands)

    VANDOORNE, H; DEVRINGER, T

    The survival of Candida albicans and Aspergillus niger in O/W creams with different types and concentrations of parabens was studied. The survival was not only dependent on the type and concentration of the preservative, but also on the size of the inoculum. The results are relevant for future

  3. The weak acid preservative sorbic acid inhibits conidial germination and mycelial growth of Aspergillus niger through intracellular acidification

    NARCIS (Netherlands)

    Plumridge, A.; Hesse, S.J.A.; Watson, A.J.; Lowe, K.C.; Stratford, M.; Archer, D.B.

    2004-01-01

    The growth of the filamentous fungus Aspergillus niger, a common food spoilage organism, is inhibited by the weak acid preservative sorbic acid (trans-trans-2,4-hexadienoic acid). Conidia inoculated at 105/ml of medium showed a sorbic acid MIC of 4.5 mM at pH 4.0, whereas the MIC for the amount of

  4. Selection and characterisation of a xylitol-derepressed Aspergillus niger mutant that is apparently impaired in xylitol transport

    NARCIS (Netherlands)

    Vondervoort, van de P.J.I.; Groot, de M.J.L.; Ruijter, G.J.G.; Visser, J.

    2006-01-01

    Aspergillus niger is known for its biotechnological applications, such as the use of xylanase enzyme for the degradation of hemicellulose. Depending on culture conditions, several polyols may also be accumulated, such as xylitol during D-xylose oxidation. Also during industrial fermentation of

  5. Comparative genomics of citric-acid-producing Aspergillus niger ATCC 1015 versus enzyme-producing CBS 513.88

    DEFF Research Database (Denmark)

    Andersen, Mikael Rørdam; Salazar, Margarita Pena; Schaap, Peter J.

    2011-01-01

    The filamentous fungus Aspergillus niger exhibits great diversity in its phenotype. It is found globally, both as marine and terrestrial strains, produces both organic acids and hydrolytic enzymes in high amounts, and some isolates exhibit pathogenicity. Although the genome of an industrial enzym...

  6. Improved mannan-degrading enzymes' production by Aspergillus niger through medium optimization.

    Science.gov (United States)

    Mohamad, Siti Norita; Ramanan, Ramakrishnan Nagasundara; Mohamad, Rosfarizan; Ariff, Arbakariya B

    2011-02-28

    The effect of different carbon and nitrogen sources on the production of mannan-degrading enzymes, focussing on β-mannanase, by Aspergillus niger was investigated using shake flask culture. The β-mannanase activity obtained during growth of A. niger on guar gum (GG, 1495 nkat mL(-1)) was much higher than those observed on other carbon substrates, locust bean gum (1148 nkat mL(-1)), α-cellulose (10.7 nkat mL(-1)), glucose (8.8 nkat mL(-1)) and carboxymethylcellulose (4.6 nkat mL(-1)). For fermentation using GG as a carbon source, bacteriological peptone gave the highest β-mannanase activity (1744 nkat mL(-1)) followed by peptone from meat (1168 nkat mL(-1)), yeast extract (817 nkat mL(-1)), ammonium sulphate (241 nkat mL(-1)), ammonium nitrate (113 nkat mL(-1)) and ammonium chloride (99 nkat mL(-1)) when used as a nitrogen source. The composition of bacteriological peptone and initial pH of the medium were further optimized using response surface methodology (RSM). Medium consisted of 21.3 g L(-1) GG and 57 g L(-1) peptone with initial culture pH of 5.5 was optimum for β-mannanase production (2063 nkat mL(-1)) by A. niger. The β-mannanase production obtained in this study using A. niger was significantly higher than those reported in the literature. Copyright © 2010 Elsevier B.V. All rights reserved.

  7. Morphology engineering - Osmolality and its effect on Aspergillus niger morphology and productivity

    Science.gov (United States)

    2011-01-01

    Background The filamentous fungus Aspergillus niger is a widely used strain in a broad range of industrial processes from food to pharmaceutical industry. One of the most intriguing and often uncontrollable characteristics of this filamentous organism is its complex morphology, ranging from dense spherical pellets to viscous mycelia depending on culture conditions. Optimal productivity correlates strongly with a specific morphological form, thus making high demands on process control. Results In about 50 2L stirred tank cultivations the influence of osmolality on A. niger morphology and productivity was investigated. The specific productivity of fructofuranosidase producing strain A. niger SKAn 1015 could be increased notably from 0.5 to 9 U mg-1 h-1 around eighteen fold, by increasing the culture broth osmolality by addition of sodium chloride. The specific productivity of glucoamylase producing strain A. niger AB1.13, could be elevated using the same procedure. An optimal producing osmolality was shown to exist well over the standard osmolality at about 3.2 osmol kg-1 depending on the strain. Fungal morphology of all cultivations was examined by microscope and characterized by digital image analysis. Particle shape parameters were combined to a dimensionless Morphology number, which enabled a comprehensive characterization of fungal morphology correlating closely with productivity. A novel method for determination of germination time in submerged cultivations by laser diffraction, introduced in this study, revealed a decelerated germination process with increasing osmolality. Conclusions Through the introduction of the versatile Morphology number, this study provides the means for a desirable characterization of fungal morphology and demonstrates its relation to productivity. Furthermore, osmolality as a fairly new parameter in process engineering is introduced and found to affect fungal morphology and productivity. Osmolality might provide an auspicious and

  8. Exploiting proteomic data for genome annotation and gene model validation in Aspergillus niger

    Directory of Open Access Journals (Sweden)

    Grigoriev Igor V

    2009-02-01

    Full Text Available Abstract Background Proteomic data is a potentially rich, but arguably unexploited, data source for genome annotation. Peptide identifications from tandem mass spectrometry provide prima facie evidence for gene predictions and can discriminate over a set of candidate gene models. Here we apply this to the recently sequenced Aspergillus niger fungal genome from the Joint Genome Institutes (JGI and another predicted protein set from another A.niger sequence. Tandem mass spectra (MS/MS were acquired from 1d gel electrophoresis bands and searched against all available gene models using Average Peptide Scoring (APS and reverse database searching to produce confident identifications at an acceptable false discovery rate (FDR. Results 405 identified peptide sequences were mapped to 214 different A.niger genomic loci to which 4093 predicted gene models clustered, 2872 of which contained the mapped peptides. Interestingly, 13 (6% of these loci either had no preferred predicted gene model or the genome annotators' chosen "best" model for that genomic locus was not found to be the most parsimonious match to the identified peptides. The peptides identified also boosted confidence in predicted gene structures spanning 54 introns from different gene models. Conclusion This work highlights the potential of integrating experimental proteomics data into genomic annotation pipelines much as expressed sequence tag (EST data has been. A comparison of the published genome from another strain of A.niger sequenced by DSM showed that a number of the gene models or proteins with proteomics evidence did not occur in both genomes, further highlighting the utility of the method.

  9. Cytosolic streaming in vegetative mycelium and aerial structures of Aspergillus niger.

    Science.gov (United States)

    Bleichrodt, R; Vinck, A; Krijgsheld, P; van Leeuwen, M R; Dijksterhuis, J; Wösten, H A B

    2013-03-15

    Aspergillus niger forms aerial hyphae and conidiophores after a period of vegetative growth. The hyphae within the mycelium of A. niger are divided by septa. The central pore in these septa allows for cytoplasmic streaming. Here, we studied inter- and intra-compartmental streaming of the reporter protein GFP in A. niger. Expression of the gene encoding nuclear targeted GFP from the gpdA or glaA promoter resulted in strong fluorescence of nuclei within the vegetative hyphae and weak fluorescence in nuclei within the aerial structures. These data and nuclear run on experiments showed that gpdA and glaA are higher expressed in the vegetative mycelium when compared to aerial hyphae, conidiophores and conidia. Notably, gpdA or glaA driven expression of the gene encoding cytosolic GFP resulted in strongly fluorescent vegetative hyphae and aerial structures. Apparently, GFP streams from vegetative hyphae into aerial structures. This was confirmed by monitoring fluorescence of photo-activatable GFP (PA-GFP). In contrast, PA-GFP did not stream from aerial structures to vegetative hyphae. Streaming of PA-GFP within vegetative hyphae or within aerial structures of A. niger occurred at a rate of 10-15 μm s(-1). Taken together, these results not only show that GFP streams from the vegetative mycelium to aerial structures but it also indicates that its encoding RNA is not streaming. Absence of RNA streaming would explain why distinct RNA profiles were found in aerial structures and the vegetative mycelium by nuclear run on analysis and micro-array analysis.

  10. An artificially constructed Syngonium podophyllum-Aspergillus niger combinate system for removal of uranium from wastewater.

    Science.gov (United States)

    He, Jia-dong; Wang, Yong-dong; Hu, Nan; Ding, Dexin; Sun, Jing; Deng, Qin-wen; Li, Chang-wu; Xu, Fei

    2015-12-01

    Aspergillus niger was inoculated to the roots of five plants, and the Syngonium podophyllum-A. niger combinate system (SPANCS) was found to be the most effective in removing uranium from hydroponic liquid with initial uranium concentration of 5 mg L(-1). Furthermore, the hydroponic experiments on the removal of uranium from the hydroponic liquids with initial uranium concentrations of 0.5, 1.0, and 3.0 mg L(-1) by the SPANCS were conducted, the inhibitory effect of A. niger on the growth of S. podophyllum in the SPANCS was studied, the accumulation characteristics of uranium by S. podophyllum in the SPANCS were analyzed, and the Fourier transform infrared (FT-IR) and extended X-ray absorption fine structure (EXAFS) spectra were measured. The results show that the removal of uranium by the SPANCS from the hydroponic liquids with initial uranium concentrations of 0.5, 1.0, and 3.0 mg L(-1) reached 98.20, 97.90, and 98.50%, respectively, after 37 days of accumulation of uranium; that the uranium concentrations in the hydroponic liquids decreased to 0.009, 0.021, and 0.045 mg L(-1), respectively, which are lower than the stipulated concentration for discharge of 0.050 mg L(-1) by the People's Republic of China; that A. niger helped to generate more groups in the root of S. podophyllum which can improve the complexing capability of S. podophyllum for uranium; and that the uranium accumulated in the root of S. podophyllum was in the form of phosphate uranyl and carboxylic uranyl.

  11. Morphology engineering--osmolality and its effect on Aspergillus niger morphology and productivity.

    Science.gov (United States)

    Wucherpfennig, Thomas; Hestler, Timo; Krull, Rainer

    2011-07-29

    The filamentous fungus Aspergillus niger is a widely used strain in a broad range of industrial processes from food to pharmaceutical industry. One of the most intriguing and often uncontrollable characteristics of this filamentous organism is its complex morphology, ranging from dense spherical pellets to viscous mycelia depending on culture conditions. Optimal productivity correlates strongly with a specific morphological form, thus making high demands on process control. In about 50 2L stirred tank cultivations the influence of osmolality on A. niger morphology and productivity was investigated. The specific productivity of fructofuranosidase producing strain A. niger SKAn 1015 could be increased notably from 0.5 to 9 U mg(-1) h(-1) around eighteen fold, by increasing the culture broth osmolality by addition of sodium chloride. The specific productivity of glucoamylase producing strain A. niger AB1.13, could be elevated using the same procedure. An optimal producing osmolality was shown to exist well over the standard osmolality at about 3.2 osmol kg(-1) depending on the strain. Fungal morphology of all cultivations was examined by microscope and characterized by digital image analysis. Particle shape parameters were combined to a dimensionless Morphology number, which enabled a comprehensive characterization of fungal morphology correlating closely with productivity. A novel method for determination of germination time in submerged cultivations by laser diffraction, introduced in this study, revealed a decelerated germination process with increasing osmolality. Through the introduction of the versatile Morphology number, this study provides the means for a desirable characterization of fungal morphology and demonstrates its relation to productivity. Furthermore, osmolality as a fairly new parameter in process engineering is introduced and found to affect fungal morphology and productivity. Osmolality might provide an auspicious and reliable approach to increase

  12. Treatment of poplar alkaline peroxide mechanical pulping (APMP) effluent with Aspergillus niger.

    Science.gov (United States)

    Liu, Tingzhi; Hu, Huiren; He, Zhibin; Ni, Yonghao

    2011-08-01

    Although the moderate load (COD of 5000-10,000 mg/L) and biodegradability of the APMP pulping effluent should allow biological treatment, toxic compounds in the effluent can interfere with this type of treatment. Studies were conducted to determine if treatment of the effluent with Aspergillus niger S13 was feasible. Under the optimized conditions (3% inoculum, pH 6, shaking at 160 rpm, 60-72 h, and 30°C), this fungus was able to remove about 97% of the methyl tertiary butyl ether (MTBE) extractives, and 60%, 77% and 43% of the chemical oxygen demand, turbidity and color even without a pre-flocculation step. These results are of practical interest in China because the APMP process has become popular, and efficient and cost-effective effluent treatment technologies are in high demand. Copyright © 2011 Elsevier Ltd. All rights reserved.

  13. Metabolic flux analysis for optimizing the specific growth rate of recombinant Aspergillus niger.

    Science.gov (United States)

    Gheshlaghi, R; Scharer, J M; Moo-Young, M; Douglas, P L

    2007-11-01

    A comprehensive metabolic network comprising three intracellular compartments (cytoplasm, mitochondrion and peroxisome) was developed for Aspergillus niger. The metabolic flux network includes carbohydrate and amino acid metabolism in both anabolic and catabolic reactions. Linear programming was used for the optimization of the specific growth rates in combination with 37 measured input and output fluxes of the key metabolites to evaluate corresponding intracellular flux distributions throughout the batch fermentations. Logarithmic sensitivity analysis revealed that the addition of proline, alanine and glutamate benefited growth in defined media. The experimental observations and flux analysis showed that tyrosine was a potential candidate for biomass production improvement. Model predictions was verified by conducting batch and fed-batch fermentations and it was found that the addition of the four amino acids according to the predetermined schedule resulted in a 44 and 41% improvements in biomass and recombinant protein productions, respectively.

  14. Influence of acid phosphatase activity on the saccharification of potato maltodextrins by Aspergillus niger glucoamylase

    Energy Technology Data Exchange (ETDEWEB)

    Zyla, K. (Akademia Rolnicza, Cracow (Poland). Dept. of Biotechnology)

    1990-01-01

    A preparation of Aspergillus niger acid phosphatase, which had the temperature optimum 60deg C, pH optimum 1.8-3.0; good stability at pH 4-5, the ability to hydrolyze glucose-6-phosphate at a high rate, and substantial lack of glucogenic activities, was used simultaneously with a glucoamylase in order to learn its influence on the saccharification of potato maltodextrins. The addition of the acid phosphatase activity in amounts that gave the 50 fold increase, as compared to phosphatase activity which naturally occurs in the gluocoamylase (GA) preparation 'AMG-200', was found to influence on the DE level, mainly at the high substrate concentration (40% d.s.) and low glucoamylase dosage (60-100 GAU/kg d.s.). It may also be possible, when using the acid phosphatase addition, to shorten the saccharification time. (orig.).

  15. Refinement of the crystal structures of biomimetic weddellites produced by microscopic fungus Aspergillus niger

    Science.gov (United States)

    Rusakov, A. V.; Frank-Kamenetskaya, O. V.; Gurzhiy, V. V.; Zelenskaya, M. S.; Izatulina, A. R.; Sazanova, K. V.

    2014-05-01

    The single-crystal structures of four biomimetic weddellites CaC2O4 · (2 + x)H2O with different contents of zeolitic water ( x = 0.10-0.24 formula units) produced by the microscopic fungus Aspergillus niger were refined from X-ray diffraction data ( R = 0.029-0.038). The effect of zeolitic water content on the structural stability of weddellite was analyzed. The parameter a was shown to increase with increasing x due to the increase in the distance between water molecules along this direction. The water content and structural parameters of the synthesized weddellites are similar to those of weddellites from biofilms and kidney stones.

  16. Structure-activity relationship of citrus polymethoxylated flavones and their inhibitory effects on Aspergillus niger.

    Science.gov (United States)

    Liu, Li; Xu, Xiaoyun; Cheng, Dan; Yao, Xiaolin; Pan, Siyi

    2012-05-02

    Citrus peels are rich in polymethoxylated flavones (PMFs) and are potential sources of natural preservatives. Six PMFs extracts, isolated and purified from the peels of three mandarins (Citrus reticulata) and three sweet oranges (Citrus sinensis), were identified and quantitated. Their inhibitory effects on Aspergillus niger were evaluated using a microbroth dilution assay. The Red tangerine variety exhibited the greatest antifungal activity (MIC = 0.2 mg/mL), while Jincheng showed the lowest activity (MIC = 1.8 mg/mL). An analysis of principal components was applied to the results in order to elucidate the structure-activity relationships of the citrus PMFs. The structure-activity relationship analysis revealed that, for good inhibitory effect, the 5-OH, 3-OCH₃, and 8-OCH₃ functionalities were essential, while the presence of 3-OH and 3'-OCH₃ greatly reduced inhibition. The findings of this study provide important information for the exploitation and utilization of citrus PMFs as natural biopreservatives.

  17. Kinetics of the hydrolysis of di- and trisaccharides with Aspergillus niger glucoamylases I and II.

    Science.gov (United States)

    Meagher, M M; Reilly, P J

    1989-08-20

    Near-homogeneous forms of glucoamylases I and II, previously purified from an industrial Aspergillus niger preparation, were used to hydrolyze a number of di- and trisaccharides linked by alpha-D-glucosidic bonds. Maximum rates and Michaelis constants were obtained at various temperatures and pH values with glucoamylase I for the disaccharides beta,alpha-trehalose, kojibiose, nigerose, maltose, and isomaltose and the trisaccharides panose and iso-maltotriose, and with glucoamylase II for maltose, maltotriose, and isomaltotriose. Maximum rates were highest and energies of activation were lowest for maltose, maltotriose, and panose, the only three substrates containing alpha-D-(1, 4)-glucosidic bonds. Michaelis constants were lowest and standard heats of binding were most negative for maltose and maltotriose. The variation of maximum rates and Michaelis constants with varying pH values suggested that two carboxyl groups were involved in substrate binding.

  18. Biotransformations of imbricatolic acid by Aspergillus niger and Rhizopus nigricans cultures.

    Science.gov (United States)

    Schmeda-Hirschmann, Guillermo; Aranda, Carlos; Kurina, Marcela; Rodríguez, Jaime A; Theoduloz, Cristina

    2007-05-21

    Microbial transformation of imbricatolic acid (1) by Aspergillus niger afforded 1alpha-hydroxyimbricatolic acid (2), while transformation with Rhizopus nigricans yielded 15-hydroxy-8,17-epoxylabdan-19-oic acid (3). When the diterpene 1 was added to a Cunninghamella echinulata culture, the main products were the microbial metabolites mycophenolic acid (4) and its 3-hydroxy derivative 5. All the structures were elucidated by spectroscopic methods. The cytotoxicity of these compounds towards human lung fibroblasts and AGS cells was assessed. While 4 and 5 showed low cytotoxicity, with IC50 values > 1000 microM against AGS cells and fibroblasts, 1alpha-hydroxyimbricatolic acid (2) presented moderate toxicity towards these targets, with IC50 values of 307 and 631 microM, respectively. The structure of 2 is presented for the first time.

  19. Biotransformations of Imbricatolic Acid by Aspergillus niger and Rhizopus nigricans Cultures

    Directory of Open Access Journals (Sweden)

    Cristina Theoduloz

    2007-05-01

    Full Text Available Microbial transformation of imbricatolic acid (1 by Aspergillus niger afforded 1α-hydroxyimbricatolic acid (2, while transformation with Rhizopus nigricans yielded 15-hydroxy-8,17-epoxylabdan-19-oic acid (3. When the diterpene 1 was added to a Cunninghamella echinulata culture, the main products were the microbial metabolites mycophenolic acid (4 and its 3-hydroxy derivative 5. All the structures were elucidated by spectroscopic methods. The cytotoxicity of these compounds towards human lung fibroblasts and AGS cells was assessed. While 4 and 5 showed low cytotoxicity, with IC50 values > 1000 μM against AGS cells and fibroblasts, 1α-hydroxyimbricatolic acid (2 presented moderate toxicity towards these targets, with IC50 values of 307 and 631 μM, respectively. The structure of 2 is presented for the first time.

  20. Transcription factors as targets for improving Aspergillus niger as cell factory

    DEFF Research Database (Denmark)

    Poulsen, Lars; Bruno, K.S.; Thykær, Jette

    Altering fluxes for overcoming metabolic bottlenecks have traditionally been approached by genetic engineering of a single or few metabolic genes. This strategy struggles to overcome the subjacent regulation thus the outcome has frequently shown to be of limited success. Transcription factors have...... the potential of controlling several fluxes in an organism, hence manipulating expression of these proteins can provide an alternative tool for overcoming metabolic bottlenecks. This approach has previously been demonstrated in yeast with great success for production of ethanol (Schuurmans et al., 2008......). In the present study the effect of modulation of transcription factors in Aspergillus niger, which is an industrially important micro-organism used in various processes including organic acid and enzyme production, was investigated. The strategy described in this work focuses on regulation connected to p...

  1. Improving Aspergillus niger as a production host through manipulation of pH responding transcription factors

    DEFF Research Database (Denmark)

    Poulsen, Lars; Bruno, K.S.; Thykær, Jette

    Altering fluxes for overcoming metabolic bottlenecks have traditionally been approached by genetic engineering of a single or few metabolic genes. This strategy struggles to overcome the subjacent regulation thus the outcome has frequently shown to be of limited success. Transcription factors have...... the potential of controlling several fluxes in an organism, hence manipulating expression of these proteins can provide an alternative tool for overcoming metabolic bottlenecks. This approach has previously been demonstrated in yeast with great success for production of ethanol (Schuurmans et al., 2008......). In the present study the effect of modulation of transcription factors in Aspergillus niger, which is an industrially important micro-organism used in various processes including organic acid and enzyme production, was investigated. The strategy described in this work focuses on regulation connected to p...

  2. Microbial carbonylation and hydroxylation of 20(R)-panaxadiol by Aspergillus niger.

    Science.gov (United States)

    Yan, Bin; Chen, Zhihua; Zhai, Xuguang; Yin, Guibo; Ai, Yafei; Chen, Guangtong

    2018-04-01

    20(R)-panaxadiol (PD) was metabolised by the fungus Aspergillus niger AS 3.3926 to its C-3 carbonylated metabolite and five other hydroxylated metabolites (1-6). Their structures were elucidated as 3-oxo-20(R)-panaxadiol (1), 3-oxo-7β-hydroxyl- 20(R)-panaxadiol (2), 3-oxo-7β,23α-dihydroxyl-20(R)-panaxadiol (3), 3,12-dioxo- 7β,23β-dihydroxyl-20(R)-panaxadiol (4), 3-oxo-1α,7β-dihydroxyl-20(R)-panaxadiol (5) and 3-oxo-7β,15β-dihydroxyl-20(R)-panaxadiol (6) by spectroscopic analysis. Among them, compounds 2-6 were new compounds. Pharmacological studies revealed that compound 6 exhibited significant anti-hepatic fibrosis activity.

  3. Optimization of Gallic Acid Production from Terminalia Chebula by Aspergillus niger

    Directory of Open Access Journals (Sweden)

    N. Lokeswari

    2007-01-01

    Full Text Available A method for producing gallic acid by microbiological hydrolysis of the tannins of myrobalan seed powder is described in the present work. Hydrolysis of gallotannins of the substrate to gallic acid by Aspergillus niger MTCC 282 was studied. A simple extraction procedure is used. Fungal mycelia pre-induced with 5 g/L gallotannin was used as inoculums. Optimal conditions of production were determined using various parameters including gallotannin concentration, nutritional source and metal ions are determined. Gallotannin is hydrolyzed with acid, and gallic acid in the hydrolyses is then assayed using rhodanine. This method is very specific: no interferences from other plant phenolics, including ellagic acid and condensed tannin, have been observed. The yield of gallic acid with respect to gallotannins present in the substrate is estimated. Yields of gallic acid are about 74% with respect to gallotannin concentration, which suggests that this method is exploitable industrially for the manufacturing trimethoprim drug.

  4. Immobilized Aspergillus niger Lipase with SiO2 Nanoparticles in Sol-Gel Materials

    Directory of Open Access Journals (Sweden)

    Li Xu

    2016-09-01

    Full Text Available Lipase from Aspergillus niger was “doubly immobilized” with SiO2 nanoparticles in sol-gel powders prepared via the base-catalyzed polymerization of tetramethoxysilane (TMOS and methyltreimethoxysilane (MTMS. The hydrolytic activity of the immobilized lipase was measured using the p-nitrophenyl palmitate hydrolysis method. The results showed that the optimum preparation conditions for the gels were made using a MTMS/TMOS molar ratio of 5, 60 mg of SiO2 nanoparticles, a water/silane molar ratio of 12, 120 mg of enzyme supply, and 120 μL of PEG400. Under the optimal conditions, the immobilized lipase retained 92% of the loading protein and 94% of the total enzyme activity. Characteristic tests indicated that the immobilized lipase exhibited much higher thermal and pH stability than its free form, which shows great potential for industrial applications.

  5. Effect of Microgravity on Fungistatic Activity of an α-Aminophosphonate Chitosan Derivative against Aspergillus niger.

    Directory of Open Access Journals (Sweden)

    Kesavan Devarayan

    Full Text Available Biocontamination within the international space station is ever increasing mainly due to human activity. Control of microorganisms such as fungi and bacteria are important to maintain the well-being of the astronauts during long-term stay in space since the immune functions of astronauts are compromised under microgravity. For the first time control of the growth of an opportunistic pathogen, Aspergillus niger, under microgravity is studied in the presence of α-aminophosphonate chitosan. A low-shear modelled microgravity was used to mimic the conditions similar to space. The results indicated that the α-aminophosphonate chitosan inhibited the fungal growth significantly under microgravity. In addition, the inhibition mechanism of the modified chitosan was studied by UV-Visible spectroscopy and cyclic voltammetry. This work highlighted the role of a bio-based chitosan derivative to act as a disinfectant in space stations to remove fungal contaminants.

  6. OPTIMIZATION OF EXTRACELLULAR TANNASE PRODUCTION BY ASPERGILLUS NIGER VAN TIEGHEM USING RESPONSE SURFACE METHODOLOGY

    Directory of Open Access Journals (Sweden)

    Hamada Abou-Bakr

    2013-12-01

    Full Text Available Response surface methodology (RSM was used to optimize the production of tannase by a newly isolate of Aspergillus niger Van Tieghem using rotatable central composite design (RCCD. This statistical optimization process was carried out involving four of quantitative growth parameters (variables, namely tannic acid concentration, nitrogen source concentration, initial pH of the medium and inoculum size. A mathematical model expressing the production process of tannase by submerged fermentation (SmF technique was generated statistically in the form of a second order polynomial equation. The model indicated the presence of significant linear, quadratic and interaction effects of the studied variables on tannase production by the fungal isolate. The results showed maximum tannase production (580 U/50 ml medium at 2% tannic acid, 4 g/l sodium nitrate, pH 4 and inoculum size of 5×107 spores/50 ml medium, which was also verified by experimental data.

  7. Improvement of xylanase production by a parasexual cross between Aspergillus niger strains

    Directory of Open Access Journals (Sweden)

    Octavio Loera

    2003-03-01

    Full Text Available A diploid strain (D4 isolated via parasexual recombination between two Aspergillus niger xylanase overproducing mutants was characterised in terms of enzyme production and catabolite repression by glucose. This strain increased xylanase production (607 nkat/ml, which was nearly 100% higher than titers achieved by the wild type strain (305 nkat/ml and 28% higher than the best mutant used to induce parasexual cycle. Diploid D4 was also less sensitive to carbon catabolite repression by glucose, since xylanolytic activity was detected under conditions normally repressing production by the wild type strain. No decrease in maximal xylanase levels was observed in the presence of glucose for diploid D4.Um cepa diplóide (D4 isolada por combinação parasexual entre dois Aspergillus niger, mutantes superprodutores de xylanase foi caracterizado através da produção de (607 nkat/ml e repressão catabólica por glicose. Essa cepa aumenta a produção de xylanase em mais de 100% em comparação com uma cepa selvagem (305 nkat/ml e 28% superior do que o melhor mutante usado para induzir o ciclo parasexual. A cepa diplóide D4 foi também menos sensível a repressão catabólica pela glicose, sendo que a atividade xylanolitica foi detectada sob condições normalmente de produção repressiva pela cepa selvagem. Não foi observado um decréscimo na produção máxima de xylanase em presença de glicose para o diplóide D4.

  8. Oxalic acid production by citric acid-producing Aspergillus niger overexpressing the oxaloacetate hydrolase gene oahA.

    Science.gov (United States)

    Kobayashi, Keiichi; Hattori, Takasumi; Honda, Yuki; Kirimura, Kohtaro

    2014-05-01

    The filamentous fungus Aspergillus niger is used worldwide in the industrial production of citric acid. However, under specific cultivation conditions, citric acid-producing strains of A. niger accumulate oxalic acid as a by-product. Oxalic acid is used as a chelator, detergent, or tanning agent. Here, we sought to develop oxalic acid hyperproducers using A. niger as a host. To generate oxalic acid hyperproducers by metabolic engineering, transformants overexpressing the oahA gene, encoding oxaloacetate hydrolase (OAH; EC 3.7.1.1), were constructed in citric acid-producing A. niger WU-2223L as a host. The oxalic acid production capacity of this strain was examined by cultivation of EOAH-1 under conditions appropriate for oxalic acid production with 30 g/l glucose as a carbon source. Under all the cultivation conditions tested, the amount of oxalic acid produced by EOAH-1, a representative oahA-overexpressing transformant, exceeded that produced by A. niger WU-2223L. A. niger WU-2223L and EOAH-1 produced 15.6 and 28.9 g/l oxalic acid, respectively, during the 12-day cultivation period. The yield of oxalic acid for EOAH-1 was 64.2 % of the maximum theoretical yield. Our method for oxalic acid production gave the highest yield of any study reported to date. Therefore, we succeeded in generating oxalic acid hyperproducers by overexpressing a single gene, i.e., oahA, in citric acid-producing A. niger as a host.

  9. Data on the presence or absence of genes encoding essential proteins for ochratoxin and fumonisin biosynthesis in Aspergillus niger and Aspergillus welwitschiae

    Directory of Open Access Journals (Sweden)

    Fernanda Pelisson Massi

    2016-06-01

    Full Text Available We present the multiplex PCR data for the presence/absence of genes involved in OTA and FB2 biosynthesis in Aspergillus niger/Aspergillus welwitschiae strains isolated from different food substrates in Brazil. Among the 175 strains analyzed, four mPCR profiles were found: Profile 1 (17% highlights strains harboring in their genome the pks, radH and the fum8 genes. Profile 2 (3.5% highlights strains harboring genes involved in OTA biosynthesis i.e. radH and pks. Profile 3 (51.5% highlights strains harboring the fum8 gene. Profile 4 (28% highlights strains not carrying the genes studied herein. This research content is supplemental to our original research article, “Prospecting for the incidence of genes involved in ochratoxin and fumonisin biosynthesis in Brazilian strains of A. niger and A. welwitschiae” [1].

  10. Spores of Aspergillus niger as reservoir of glucose oxidase synthesized during solid‐state fermentation and their use as catalyst in gluconic acid production

    National Research Council Canada - National Science Library

    Ramachandran, S; Fontanille, P; Pandey, A; Larroche, C

    2007-01-01

    Aims:  To exploit conidiospores of Aspergillus niger as a vector for glucose oxidase extraction from solid media, and their direct use as biocatalyst in the bioconversion of glucose to gluconic acid...

  11. Oxalic acid production by Aspergillus niger: an oxalate-non-producing mutant produces citric acid at pH 5 and in the presence of manganese

    National Research Council Canada - National Science Library

    Ruijter, George J. G; van de Vondervoort, Peter J. I; Visser, Jaap

    1999-01-01

    ... for correspondence: Jaap Visser. Tel: +31 317 484439. Fax: +31 317 484011. e-mail: office{at}algemeen.mgim.wau.nl The external pH appeared to be the main factor governing oxalic acid production by Aspergillus niger...

  12. Optimization of the production of Aspergillus niger α-glucosidase expressed in Pichia pastoris.

    Science.gov (United States)

    Liu, Xu; Wu, Dan; Wu, Jing; Chen, Jian

    2013-03-01

    The α-glucosidase (AGL) from Aspergillus niger has been applied to produce isomaltooligosaccharides. In the present study, various factors which affect the yield of recombinant AGL, produced by engineered Pichia pastoris, were investigated. The expression level reached 5.5 U ml(-1) in bioreactor after optimization of parameters of initial induction cell density, induction temperature and methanol concentration. In addition, it was found that coexpression of protein disulfide isomerase (PDI) inhibited the growth of the engineered P. pastoris strains and had an adverse effect on the production of AGL, while codon optimization of native A. niger α-glucosidase encoding gene (aglu) resulted in a significant enhancement of enzyme production, which reached 10.1 U ml(-1). We believe that yield of AGL is increased by codon optimization as a result of enhanced translation efficiency as well as more stable mRNA secondary structure. In contrast, PDI coexpression under the control of alcohol oxidase promoter (PAOX1) seems to be less efficient in helping disulfide bond formation in AGL while probably induce unfolded protein response, which further leads to cell apoptosis and increased protein degradation.

  13. Bioprocessing of citrus waste peel for induced pectinase production by Aspergillus niger; its purification and characterization

    Directory of Open Access Journals (Sweden)

    Ishtiaq Ahmed

    2016-04-01

    Full Text Available Agro-industrial residues are primarily composed of complex polysaccharides that strengthen microbial growth for the production of industrially important enzymes. Pectinases are one of the most widely disseminated enzymes in bacteria, fungi and plants. Czapeck media supplemented with orange waste peel as carbon source under submerged fermentation process Aspergillus niger presenting the preeminent enzymatic production. On partial optimization culture showed the maximum enzyme yield (117.1 ± 3.4 μM/mL/min at 30 °C in an orange waste peel medium having pH 5.5 and substrate concentration (4% after 5th day of fermentation. The produced enzyme was purified by ammonium sulfate precipitation and ion exchange chromatography. A purification fold of 5.59 with specific activity and % recovery of 97.2 U/mg and 12.96% was achieved respectively after gel filtration chromatographic technique. The molecular weight of purified pectinase from A. niger was 30 kDa evidenced by SDS-PAGE. Pectinase activity profile showed purified enzyme was optimally active at pH = 7 and 55 °C. The maximum production of pectinase in the presence of cheaper substrate at low concentration makes the enzyme useful in industrial sectors especially for textile and juice industry.

  14. Spatial differentiation of gene expression in Aspergillus niger colony grown for sugar beet pulp utilization.

    Science.gov (United States)

    Benoit, Isabelle; Zhou, Miaomiao; Vivas Duarte, Alexandra; Downes, Damien J; Todd, Richard B; Kloezen, Wendy; Post, Harm; Heck, Albert J R; Maarten Altelaar, A F; de Vries, Ronald P

    2015-08-28

    Degradation of plant biomass to fermentable sugars is of critical importance for the use of plant materials for biofuels. Filamentous fungi are ubiquitous organisms and major plant biomass degraders. Single colonies of some fungal species can colonize massive areas as large as five soccer stadia. During growth, the mycelium encounters heterogeneous carbon sources. Here we assessed whether substrate heterogeneity is a major determinant of spatial gene expression in colonies of Aspergillus niger. We analyzed whole-genome gene expression in five concentric zones of 5-day-old colonies utilizing sugar beet pulp as a complex carbon source. Growth, protein production and secretion occurred throughout the colony. Genes involved in carbon catabolism were expressed uniformly from the centre to the periphery whereas genes encoding plant biomass degrading enzymes and nitrate utilization were expressed differentially across the colony. A combined adaptive response of carbon-catabolism and enzyme production to locally available monosaccharides was observed. Finally, our results demonstrate that A. niger employs different enzymatic tools to adapt its metabolism as it colonizes complex environments.

  15. Bioleaching of valuable metals from spent lithium-ion mobile phone batteries using Aspergillus niger

    Science.gov (United States)

    Horeh, N. Bahaloo; Mousavi, S. M.; Shojaosadati, S. A.

    2016-07-01

    In this paper, a bio-hydrometallurgical route based on fungal activity of Aspergillus niger was evaluated for the detoxification and recovery of Cu, Li, Mn, Al, Co and Ni metals from spent lithium-ion phone mobile batteries under various conditions (one-step, two-step and spent medium bioleaching). The maximum recovery efficiency of 100% for Cu, 95% for Li, 70% for Mn, 65% for Al, 45% for Co, and 38% for Ni was obtained at a pulp density of 1% in spent medium bioleaching. The HPLC results indicated that citric acid in comparison with other detected organic acids (gluconic, oxalic and malic acid) had an important role in the effectiveness of bioleaching using A. niger. The results of FTIR, XRD and FE-SEM analysis of battery powder before and after bioleaching process confirmed that the fungal activities were quite effective. In addition, bioleaching achieved higher removal efficiency for heavy metals than the chemical leaching. This research demonstrated the great potential of bio-hydrometallurgical route to recover heavy metals from spent lithium-ion mobile phone batteries.

  16. Heavy metals extraction from municipal solid waste incineration fly ash using adapted metal tolerant Aspergillus niger.

    Science.gov (United States)

    Yang, Jie; Wang, Qunhui; Wang, Qi; Wu, Tingji

    2009-01-01

    This study focused on the adaptation of Aspergillus niger tolerating high concentration of heavy metals for bioleaching of fly ash. The Plackett-Burman design indicated that Al and Fe inhibited the growth of A. niger (AS 3.879 and AS 3.40) significantly. The single metal (Al and Fe) and multi-metals adapted AS 3.879 strain tolerated up to 3500 mg/L Al, 700 mg/L Fe, and 3208.1mg/L multi-metals, respectively. The order of metal extraction yield in two-step bioleaching of 60 and 70 g/L fly ash using Al adapted, multi-metals adapted and un-adapted AS 3.879 strains was as follows: multi-metals adapted>Al adapted>un-adapted. The multi-metals adapted strain grew with up to 70 g/L fly ash and secreted 256 mmol/L organic acids after 288 h, where 87.4% Cd, 64.8% Mn, 49.4% Zn and 45.9% Pb were dissolved. The extracted metals in TCLP test of the bioleached fly ash by multi-metals adapted strain were under the regulated levels in China.

  17. Comparison of glycosylation patterns of phytase from Aspergillus niger (A. ficuum) NRRL 3135 and recombinant phytase.

    Science.gov (United States)

    Panchal, T; Wodzinski, R J

    1998-08-01

    The glycosylation patterns of phytase (E C 3.1.3.8) from Aspergillus niger NRRL 3135 with recombinant phytase from A. niger van Tieghem were compared. The following characteristics were studied: size of the whole molecule, type of linkage (N-linked or O-linked oligosaccharide), profile (number of different type of oligosaccharides present), monosaccharide composition, their order, and configuration. The molecular weights of both glycoproteins, after deglycosylation, was approximately 55 kDa as determined by SDS-PAGE. Both glycoproteins were about 35.29% glycosylated (calculations were made on the basis of 85 kDa molecular weight before deglycosylation). Only N-linked oligosaccharides were present. When N-linked oligosaccharides were released and labeled, the same profile was obtained for both glycoproteins. Mannose residues were detected after digestion by combinations of various exoglycosidases. N-acetylneuraminic acid, galactose, and N-acetylglucosamine residues were not detected. Released mannose residues were (alpha 1-2,3,6) linked. The trisaccharide core structure was nonfucosylated for all the oligosaccharides released from both glycoproteins. The only major difference found between the two glycoproteins was the migration pattern of oligosaccharide bands on gel after digestion with alpha-mannosidases. The structure of N-linked oligosaccharides ranged from (Man)5(GlcNAc)2 to (Man)10 (GlcNAc)2.

  18. Pomelo peels as alternative substrate for extracellular pectinase production by Aspergillus niger HFM-8

    Directory of Open Access Journals (Sweden)

    Ibrahim, D.

    2013-12-01

    Full Text Available Aims: The aim of this work was to develop an effective bioprocess to enhance the pectinase production by solid-state cultures of Aspergillus niger HFM-8. Methodology and results: The pectinase production produced by A. niger HFM-8 was studied under solid state fermentation using Malaysian pomelo (Citrus grandis peel as the substrate. This local agricultural waste is rich with lignocellulolytic material including pectin acts as the inducer of pectinase production. Under optimized conditions, 5 g of 0.75 mm pomelo peel size, moisture content of 60% (v/w sterile distilled water pH 5.0, inoculums size of 1x10^4 spores/mL, cultivation temperature of room temperature (30 ± 2 °C, no mixing incurred and with the addition of 1% (w/w citrus pectin and 0.1% (w/w urea has produced pectinase production of 306.89 U/g substrate and 0.78 mg glucosamine/g substrate of fungal growth on the 8th day of cultivation. Conclusion, significance and impact of study: There was 48.82% increment in enzyme production after the improvement of parameters. It was found that pomelo peel is a suitable feedstock for pectinase production.

  19. Morphology of Filamentous Fungi: Linking Cellular Biology to Process Engineering Using Aspergillus niger

    Science.gov (United States)

    Krull, Rainer; Cordes, Christiana; Horn, Harald; Kampen, Ingo; Kwade, Arno; Neu, Thomas R.; Nörtemann, Bernd

    In various biotechnological processes, filamentous fungi, e.g. Aspergillus niger, are widely applied for the production of high value-added products due to their secretion efficiency. There is, however, a tangled relationship between the morphology of these microorganisms, the transport phenomena and the related productivity. The morphological characteristics vary between freely dispersed mycelia and distinct pellets of aggregated biomass. Hence, advantages and disadvantages for mycel or pellet cultivation have to be balanced out carefully. Due to this inadequate understanding of morphogenesis of filamentous microorganisms, fungal morphology, along with reproducibility of inocula of the same quality, is often a bottleneck of productivity in industrial production. To obtain an optimisation of the production process it is of great importance to gain a better understanding of the molecular and cell biology of these microorganisms as well as the approaches in biochemical engineering and particle technique, in particular to characterise the interactions between the growth conditions, cell morphology, spore-hyphae-interactions and product formation. Advances in particle and image analysis techniques as well as micromechanical devices and their applications to fungal cultivations have made available quantitative morphological data on filamentous cells. This chapter provides the ambitious aspects of this line of action, focussing on the control and characterisation of the morphology, the transport gradients and the approaches to understand the metabolism of filamentous fungi. Based on these data, bottlenecks in the morphogenesis of A. niger within the complex production pathways from gene to product should be identified and this may improve the production yield.

  20. Low-cost carbon sources for the production of a thermostable xylanase by Aspergillus niger

    Directory of Open Access Journals (Sweden)

    Ana Cláudia Elias Pião Benedetti

    2013-01-01

    Full Text Available A strain of the filamentous fungus Aspergillus niger was isolated and shown to possess extracellular xylanolytic activity. These enzymes have biotechnological potential and can be employed in various industries. This fungus produced its highest xylanase activity in a medium made up of 0.1% CaCO3 , 0.5% NaCl, 0.1% NH4 Cl, 0.5% corn steep liquor and 1% carbon source, at pH 8.0. A lowcost hemicellulose residue (powdered corncob proved to be an excellent inducer of the A. niger xylanolytic complex. Filtration of the crude culture medium with suspended kaolin was ideal for to clarify the extract and led to partial purification of the xylanolytic activity. The apparent molecular mass of the xylanase was about 32.3 kDa. Maximum enzyme activity occurred at pH 5.0 and 55-60ºC. Apparent Km was 10.41 ± 0.282 mg/mL and Vmax was 3.32 ± 0.053 U/mg protein, with birchwood xylan as the substrate. Activation energy was 4.55 kcal/mol and half-life of the crude enzyme at 60ºC was 30 minutes. Addition of 2% glucose to the culture medium supplemented with xylan repressed xylanase production, but in the presence of xylose the enzyme production was not affected.

  1. Evaluation of the Activities of Concentrated Crude Mannan-degrading Enzymes Produced by Aspergillus niger

    Directory of Open Access Journals (Sweden)

    Siti Norita, M.

    2010-01-01

    Full Text Available The mannan-degrading enzymes produced by Aspergillus niger were concentrated and the activities were evaluated. The optimum pH for β-mannanase, endoglucanase and α-galactosidase were obtained at pH 3.5 while pH optimum for β-mannosidase was occurred at pH 3.0. The β-mannanase, endoglucanase, β-mannosidase and α-galactosidase was stable at pH 3.5 to 7, pH 3.5 to 6.5, pH 4 to 7 and pH 3.5 to 5.0, respectively. The enzymes obtained in this study were characterized and defined as acidic proteins. The β-mannanases from A. niger had two optimum temperatures (at 50 °C and 60 °C and its half-life was 6 h and 4 h at 60 °C and 70 °C, respectively. The β-mannosidase, endoglucanase and α-galactosidase displayed optimal activity at 70 °C, 60 °C and 50 – 60 °C, respectively. The β-mannosidase had half-life of 1.5 h at 70 °C, while α-galactosidase had a half-life of 2.5 h at 60 °C and endoglucanase had a half-life of 6 h at 60 °C and 45 min at 70 °C.

  2. Optimization of growth conditions for xylanase production by Aspergillus niger in solid state fermentation.

    Science.gov (United States)

    Kavya, Venkatesh; Padmavathi, Tallapragada

    2009-01-01

    The objectives of the present study were isolation, identification and characterization of xylanase producing fungi, optimization of medium composition and cultural conditions for xylanase enzyme production using cheaper sources. The fungal strains were isolated from garden soil by serial dilution technique and Aspergillus niger was identified and isolated in pure form. In conformation screening by congo red test, based on the reddish zone of enzyme activity formation in oat spelt xylan agar plates, A. niger was selected and optimized for xylanase enzyme production in solid state fermentation using cheaper sources like wheat bran, rice bran, soya bran, ragi bran and dust. Maximum enzyme activity was observed in wheat bran (9.87 U/ml). The use of wheat bran as a major carbon source is particularly valuable because oat spelt xylan or birch wood xylan are more expensive. The effects of time course, incubation substrate, inoculum size, moisturizing agent, moisture content, temperature and volume of fermentation medium on the production of xylanase were studied. The maximum xylanase production (12.65 U/ml) was observed at optimized condition, incubation temperature of 28 degrees C after 6 days of incubation period while minimum production (9.38 U/ml) at unoptimized condition. The maximum production of enzyme was found to be in wheat bran when the volume of fermentation medium was kept as 10 g/250 ml conical flasks, with mineral solution as moisturizing agent and moisture ratio 1:0.7. Thus the present study proved that the fungal strain A. niger used is highly potential and useful for xylanase production.

  3. Dynamic Fumonisin B₂ Production by Aspergillus niger Intented Used in Food Industry in China.

    Science.gov (United States)

    Han, Xiaomin; Jiang, Hongru; Xu, Jin; Zhang, Jing; Li, Fengqin

    2017-07-09

    There are a total of 30 strains including 27 strains of Aspergillus niger intended used in Chinese food industry, two strains used as control and one strain isolated from corn for fumonisin (FB) production on 3 media. It was found that FB₂ production by A. niger was function-dependent and highly related to culture media, as well as incubation time. All strains studied were unable to produce FB₁ and FB₃. Almost all strains were found to produce FB₂ on corn, rice and wheat bran. Based on their intended use in the food industry, the higher level of FB₂ producers were strains used for saccharifying enzyme (n = 13) production, followed by organic acid (n = 6), tannase (n = 7) and β-galactosidase (n = 1) production, with the FB₂ mean level of 3553-10,270 μg/kg, 1059-12,036 μg/kg, 3-7 μg/kg and 2-4 μg/kg on corn, 5455-9241 μg/kg, 559-2190 μg/kg, 4-9 μg/kg and 6-10 μg/kg on rice, 5959-7709 μg/kg, 9491-17,339 μg/kg, 8-14 μg/kg and 120-222 μg/kg on wheat bran, respectively. Comparatively, strains of Fusarium verticillioide were capable of producing fumonins simultaneously with broader spectrum including FB₁, FB₂ and FB₃, but at a much lower level. In conclusion, it is necessary to evaluate FB₂ production by A. niger before intended use in the food processing industry.

  4. Fermentasi Etanol dari Limbah Padat Tapioka (Onggok oleh Aspergillus niger dan Zymomonas mobilis

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    RATNA SETYANINGSIH

    2004-05-01

    Full Text Available The aims of this research were to know the best concentration of onggokflour to produce maximum reduction sugar concentration by Aspergillus niger and efficiency of ethanol production from reduction sugar as the product of onggok flour saccharification by Zymomonas mobilis, and the ethanol concentration which produced. The framework of this research was cassava starch in onggok can be used as substance which yielded ethanol by fermentation. The ethanol yielded serves as an alternative substitution fuel for fossil fuel. This research was done in two stages; they were saccharification by A. niger and ethanol fermentation by Z. mobilis. At saccharification stage there were four level of onggok flour concentration:10%, 20%, 30% and 40%. The inoculum concentration of A. niger 10% (v/v and the amount of spore about 3.3x106 spore/mL, was inoculated each concentration of onggok flour, with the temperature of incubation 500C and this process taken five days. Parameters have been used were starch concentration before and after saccharification process, reduction sugar concentration, and pH conducted by each, every 24 hours. Stage of ethanol fermentation used concentration of Z. mobilis 10% (v/v, with the cell amount 5.1x10 7 cell/mL. Reduction sugar concentration from saccharification product that used 1.9352% and fermentation was done in 72 hours. Parameters were concentration of ethanol and reduction sugar each, every 24 hours and the growth of Z. mobilis cell every 24 hours. This research concluded that concentration of onggok flour 10% produced maximum reduction sugar concentration on the third day that was 1.1842%. Ethanol concentration yielded was 0.7% (v/v and efficiency of ethanol that was produced from reduction sugar from saccharification of onggok flour was 83% during 72 hours fermentation.

  5. Dynamic Fumonisin B2 Production by Aspergillus niger Intented Used in Food Industry in China

    Science.gov (United States)

    Han, Xiaomin; Jiang, Hongru; Xu, Jin; Zhang, Jing; Li, Fengqin

    2017-01-01

    There are a total of 30 strains including 27 strains of Aspergillus niger intended used in Chinese food industry, two strains used as control and one strain isolated from corn for fumonisin (FB) production on 3 media. It was found that FB2 production by A. niger was function-dependent and highly related to culture media, as well as incubation time. All strains studied were unable to produce FB1 and FB3. Almost all strains were found to produce FB2 on corn, rice and wheat bran. Based on their intended use in the food industry, the higher level of FB2 producers were strains used for saccharifying enzyme (n = 13) production, followed by organic acid (n = 6), tannase (n = 7) and β-galactosidase (n = 1) production, with the FB2 mean level of 3553–10,270 μg/kg, 1059–12,036 μg/kg, 3–7 μg/kg and 2–4 μg/kg on corn, 5455–9241 μg/kg, 559–2190 μg/kg, 4–9 μg/kg and 6–10 μg/kg on rice, 5959–7709 μg/kg, 9491–17,339 μg/kg, 8–14 μg/kg and 120–222 μg/kg on wheat bran, respectively. Comparatively, strains of Fusarium verticillioide were capable of producing fumonins simultaneously with broader spectrum including FB1, FB2 and FB3, but at a much lower level. In conclusion, it is necessary to evaluate FB2 production by A. niger before intended use in the food processing industry. PMID:28698485

  6. Isolation and NMR Characterization of Fumonisin B-2 and a New Fumonisin B-6 from Aspergillus niger

    DEFF Research Database (Denmark)

    Månsson, Maria; Klejnstrup, Marie Louise; Phipps, Richard Kerry

    2010-01-01

    A new fumonisin, fumonisin B-6 (1), has been isolated by cation-exchange and reverse-phase chromatography, together with fumonisin B-2 (2), from,stationary cultures of the fungus Aspergillus niger NRRL 326. Analysis of mass spectrometric and NMR data determined that FB6 is a positional isomer...... of FBI and iso-FB1, having hydroxyl functions at C3, C4, and C5. Analysis of the NMR data for FB2 showed very similar chemical shift values when compared to an authentic Fusarium FB2 standard, strongly indicating identical molecules despite that an absolute stereochemical assignment of FB2 from A. niger...

  7. Isolation and optimization of pectinase enzyme production one of useful industrial enzyme in Aspergillus niger, Rhizopus oryzae, Penicilium chrysogenum

    Directory of Open Access Journals (Sweden)

    akram songol

    2016-06-01

    Full Text Available Introduction: Pectinase enzyme is one of the most important industrial enzymes which isolated from a wide variety of microorganisms such as bacteria and filamentous fungi. This enzyme has been usually used in the fruit and textile industry. In this study, the isolation and optimization of pectinase-producing fungi on decaying rotten fruits were studied. Materials and methods: Isolation and screening of pectinase producing fungi performed through plate culture on pectin medium and staining with Lugol's iodine solution. The best strains were identified by ITS1, 4 sequencing as Aspergillus fumigatus, Rhizopus oryzae, Penicilium chrysogenum. The enzyme production was optimized by application of the five factorial design, each at three levels. These factors are carbon sources (whey, glucose and stevia, ammonium sulfate, manganese sulfate, temperature, and pH. Pectinase concentration was measured by the Miller method. Results: The results indicate that optimum condition for enzyme production for three fungi strains was obtained at 32 °C, pH = 6, 3g / L manganese sulfate, 2.75g / L of ammonium sulfate and 10g / L of each carbon source. The best experiment in obtaining the optimum enzyme contained 1.328 mg / ml of glucose for Aspergillus niger 1.284 and 1.039 mg / ml of whey for Rhizopus oryzae and Penicilium chrysogenum. Molecular weight of enzyme was about 40 and 37 kDa which was obtained by SDS- PAGE. Discussion and conclusion: The results indicate that three strains could grow in a wide range of carbon source, pH and temperature, which could be a good candidate for industrial application.

  8. Identification of Aspergillus (A. flavus and A. niger) Allergens and Heterogeneity of Allergic Patients' IgE Response.

    Science.gov (United States)

    Vermani, Maansi; Vijayan, Vannan Kandi; Agarwal, Mahendra Kumar

    2015-08-01

    Aspergillus species (A. flavus and A. niger) are important sources of inhalant allergens. Current diagnostic modalities employ crude Aspergillus extracts which only indicate the source to which the patient has been sensitized, without identifying the number and type of allergens in crude extracts. We report a study on the identification of major and minor allergens of the two common airborne Aspergillus species and heterogeneity of patients' IgE response to them. Skin prick tests were performed on 300 patients of bronchial asthma and/or allergic rhinitis and 20 healthy volunteers. Allergen specific IgE in patients' sera was estimated by enzyme allergosorbent test (EAST). Immunoblots were performed to identify major/minor allergens of Aspergillus extracts and to study heterogeneity of patients'IgE response to them. Positive cutaneous responses were observed in 17% and 14.7% of patients with A. flavus and A. niger extracts, respectively. Corresponding EAST positivity was 69.2% and 68.7%. In immunoblots, 5 allergenic proteins were identified in A. niger extract, major allergens being 49, 55.4 and 81.5 kDa. Twelve proteins bound patients' IgE in A. flavus extract, three being major allergens (13.3, 34 and 37 kDa). The position and slopes of EAST binding and inhibition curves obtained with individual sera varied from patient to patient. The number and molecular weight of IgE-binding proteins in both the Aspergillus extracts varied among patients. These results gave evidence of heterogeneity of patients' IgE response to major/minor Aspergillus allergens. This approach will be helpful to identify disease eliciting molecules in the individual patients (component resolved diagnosis) and may improve allergen-specific immunotherapy.

  9. AFM images of complexes between amylose and Aspergillus niger glucoamylase mutants, native and mutant starch binding domains: a model for the action of glucoamylase

    DEFF Research Database (Denmark)

    Morris, V. M.; Gunning, A. P.; Faults, C. B.

    2005-01-01

    Atomic force microscopy has been used to investigate the complexes formed between high molecular weight amylose chains and Aspergillus niger glucoamylase mutants (E400Q and W52F), wild-type A. niger starch binding domains (SBDS), and mutant SBDs (W563K and W590K) lacking either of the two starch ...

  10. The weak-acid preservative sorbic acid is decarboxylated and detoxified by a phenylacrylic acid decarboxylase, PadA1, in the spoilage mold Aspergillus niger.

    Science.gov (United States)

    Plumridge, Andrew; Stratford, Malcolm; Lowe, Kenneth C; Archer, David B

    2008-01-01

    Resistance to sorbic and cinnamic acids is mediated by a phenylacrylic acid decarboxylase (PadA1) in Aspergillus niger. A. niger DeltapadA1 mutants are unable to decarboxylate sorbic and cinnamic acids, and the MIC of sorbic acid required to inhibit spore germination was reduced by approximately 50% in DeltapadA1 mutants.

  11. Genomic analysis of the aconidial and high-performance protein producer, industrially relevant Aspergillus niger SH2 strain.

    Science.gov (United States)

    Yin, Chao; Wang, Bin; He, Pan; Lin, Ying; Pan, Li

    2014-05-15

    Aspergillus niger is usually regarded as a beneficial species widely used in biotechnological industry. Obtaining the genome sequence of the widely used aconidial A. niger SH2 strain is of great importance to understand its unusual production capability. In this study we assembled a high-quality genome sequence of A. niger SH2 with approximately 11,517 ORFs. Relatively high proportion of genes enriched for protein expression related FunCat items verify its efficient capacity in protein production. Furthermore, genome-wide comparative analysis between A. niger SH2 and CBS513.88 reveals insights into unique properties of A. niger SH2. A. niger SH2 lacks the gene related with the initiation of asexual sporulation (PrpA), leading to its distinct aconidial phenotype. Frame shift mutations and non-synonymous SNPs in genes of cell wall integrity signaling, β-1,3-glucan synthesis and chitin synthesis influence its cell wall development which is important for its hyphal fragmentation during industrial high-efficiency protein production. Copyright © 2014 Elsevier B.V. All rights reserved.

  12. SCREENING AND OPTIMIZATION OF CULTURE CONDITIONS FOR CELLULASE PRODUCTION BY ASPERGILLUS NIGER NSPR012 IN SUBMERGED FERMENTATION

    Directory of Open Access Journals (Sweden)

    Juliet Bamidele Akinyele

    2014-12-01

    Full Text Available This study aimed at screening of selected fungal strains and optimization of process parameters for cellulases production in submerged fermentation. Aspergillus niger NSPR012 was selected for further studies as the most potent in producing cellulase of high activity. Utilization of various agro-wastes as substitute tocarboxy methyl cellulose (CMC for cellulase production was also investigated. Among tested carbon sources, banana peels at a concentration of 5% was found to be the most effective carbon source. The cellulase production by Aspergillus niger NSPR012 in mineral salt medium attained maximum after 96 h of incubation. Maximum cellulase activity (0.466µmol/min/mL was obtained with locust beans as the best organic nitrogen source. The optimum incubation temperature and initial pH were 37°C and 5.5, respectively. With this information, banana peels could have good biotechnological potential for bio-products formation in which cellulase is one.

  13. BIOSORPSI DAN REDUKSI KROM LIMBAH PENYAMAKAN KULIT DENGAN BIOMASSA Fusarium sp DAN Aspergillus niger (Biosorpstion and Reduction of Chromium Bearing Tannery Wastewater Using The Biomass of Fusarium Sp. and Aspergillus niger

    Directory of Open Access Journals (Sweden)

    Suharjono Triatmojo

    2001-08-01

    Full Text Available ABSTRAK Tujuan penelitian ini ialah untuk membuktikan bahwa biomassa Fusarium sp dapat mereduksi Cr(VI, dan biomassa Aspergillus niger dapat digunakan untuk mengambil ion krom dari larutan. Fusarium.sp ditumbuhkan pada media cair kentang dekftosa cair, ditambah K2Cr2O7 atau sludge limbah penyamakan kulit. Selanjutnya diamati perubahan warnanya, bila terjadi perubahan warna dan oranye ke ungu atau tak berwarna maka telah terjadi reduksi krom valensi VI menjadi krom valensi Ill. Aspergillus niger ditumbuhkan pada media Potato dectrose agar (PDA padat, dipindahkan ke media cair yang bensi bakto pepton, bakto dektrose dan srukronutrien. Produksi biomassa dilakukan pada labu erlenmeyer; setelah 5 hari dipanen dan dibuat bubuk. Bubuk ini digunakan untuk mengambil krom dari larutan. Hasil penelitian menunjukkan bahwa biomassa Fusarium sp dapat digunakan untuk mengambil krom dan larutan yang.mengandung KrCrrO, atau sludge limbah penyamakan kulit. Waktu inkubasi yang lebih lama meningkatkan absorbsi krom oleh biomassa Fascrium sp. Fusarium sp mampu mereduksi Cr(VI menjadi Cr(Iii. Biomassa Aspergillus niger dapat digunakan untuk mengambil krom dari larutan. Hasil terbaik diperoleh pada konsentrasi awal 100 mg/I, pada pH 2,0, berat biomassa 0,1 g, dan waktu kontak 12 jam, yaitu 96,23% untuk Cr(II| dan96,3 % untuk Cr(VI. Fusarium sp. dan A. niger dapat digunakan sebagai bioremediator dalam penanganan limbah penyamakan kulit secara biologi.   ABSTRACT The objectives of this research was to study the biosorption and reduction of chromium bearing tannery wastewater using biomass of Fusarium sp and Aspergillus niger. Fusarium sp was used to investigate bioaccumulation and reduction of chromium in K2 Cr2O7 solution and solution containing sludge of leather tanning waste, and aspergillus niger was used to investigate biosorption of Cr(III and Cr(VI in solution. Fusarium sp was grown on sterilized potato extrose liquid medium, added with K2Cr2O7solution or sludge

  14. Statistical optimization of endo-polygalacturonase production by overproducing mutants of Aspergillus niger in solid-state fermentation

    OpenAIRE

    Siva Kiran, R.R.; Rajesh Konduri; Rao, G. H.; Madhu, G. M.

    2010-01-01

    Statistically based experimental designs were applied for endopolygalacturonase production by mutated strains of Aspergillus niger using apple pomace waste in solid-state fermentation (SSF). The Plackett-Burman design was used to search for the main factors. Urea was found to be the most significant factor (p<0.05) among these variables. To study the mutual interactions between variables and find the optimum medium, Doehlert design was performed to investigate the effect of the medium comp...

  15. Optimization of date syrup for enhancement of the production of citric acid using immobilized cells of Aspergillus niger

    OpenAIRE

    Mostafa, Yasser S.; Alamri, Saad A.

    2012-01-01

    Date syrup as an economical source of carbohydrates and immobilized Aspergillus niger J4, which was entrapped in calcium alginate pellets, were employed for enhancing the production of citric acid. Maximum production was achieved by pre-treating date syrup with 1.5% tricalcium phosphate to remove heavy metals. The production of citric acid using a pretreated medium was 38.87% higher than an untreated one that consumed sugar. The appropriate presence of nitrogen, phosphate and magnesium appear...

  16. Mycelium-bound lipase production from Aspergillus niger MYA 135, and its potential applications for the transesterification of ethanol.

    Science.gov (United States)

    Colin, Verónica Leticia; Baigorí, Mario Domingo; Pera, Licia María

    2011-06-01

    The potential biotechnological applications of both constitutive and inducible lipase sources from Aspergillus niger MYA 135 were evaluated. To this end, the effect of environmental conditions on mycelium-bound lipase production from this strain was studied, when cultured either in the absence or presence of 2% olive oil. It was previously reported that mycelium-bound lipase from Aspergillus niger MYA 135 possess high stability in reaction mixtures containing ethanol; which could be especially important for their use in biodiesel synthesis. In this connection, the performance of the lipase sources produced in the transesterification of ethanol using p-nitrophenyl palmitate as acyl donor was also explored. Under our assay conditions, hydrolytic and synthetic activity of the mycelia produced in the absence or presence of olive oil were not highly correlated. While the hydrolytic activity was strongly increased by the addition of lipid to the culture medium, the best performance in the transesterification reactions of ethanol were associated with mycelia produced in absence of olive oil. Interestingly, the supplementation of the culture medium with Fe(+3) increased the transesterification activity by 71%, as compared to the activity previously reported for this strain. Therefore, the constitutive lipase sources from Aspergillus niger MYA 135 are considered to be promising for industrial biodiesel-fuel production. Copyright © 2011 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  17. Production of antifungal chitinase by Aspergillus niger LOCK 62 and its potential role in the biological control.

    Science.gov (United States)

    Brzezinska, Maria Swiontek; Jankiewicz, Urszula

    2012-12-01

    Aspergillus niger LOCK 62 produces an antifungal chitinase. Different sources of chitin in the medium were used to test the production of the chitinase. Chitinase production was most effective when colloidal chitin and shrimp shell were used as substrates. The optimum incubation period for chitinase production by Aspergillus niger LOCK 62 was 6 days. The chitinase was purified from the culture medium by fractionation with ammonium sulfate and affinity chromatography. The molecular mass of the purified enzyme was 43 kDa. The highest activity was obtained at 40 °C for both crude and purified enzymes. The crude chitinase activity was stable during 180 min incubation at 40 °C, but purified chitinase lost about 25 % of its activity under these conditions. Optimal pH for chitinase activity was pH 6-6.5. The activity of crude and purified enzyme was stabilized by Mg(2+) and Ca(2+) ions, but inhibited by Hg(2+) and Pb(2+) ions. Chitinase isolated from Aspergillus niger LOCK 62 inhibited the growth of the fungal phytopathogens: Fusarium culmorum, Fusarium solani and Rhizoctonia solani. The growth of Botrytis cinerea, Alternaria alternata, and Fusarium oxysporum was not affected.

  18. Effect of temperature and water activity on the production of fumonisins by Aspergillus niger and different Fusarium species

    Directory of Open Access Journals (Sweden)

    Frisvad Jens C

    2009-12-01

    Full Text Available Abstract Background Fumonisins are economically important mycotoxins which until recently were considered to originate from only a few Fusarium species. However recently a putative fumonisin gene cluster was discovered in two different Aspergillus niger strains followed by detection of an actual fumonisin B2 (FB2 production in four strains of this biotechnologically important workhorse. Results In the present study, a screening of 5 A. niger strains and 25 assumed fumonisin producing Fusarium strains from 6 species, showed that all 5 A. niger strains produced FB2 and 23 of 25 Fusarium produced fumonisin B1 and other isoforms (fumonisin B2 and B3. Five A. niger and five Fusarium spp. were incubated at six different temperatures from 15-42°C on Czapek Yeast Agar +5% salt or Potato Dextrose Agar. A. niger had the highest production of FB2 at 25-30°C whereas Fusarium spp. had the maximal production of FB1 and FB2 at 20-25°C. Addition of 2.5-5% NaCl, or 10-20% sucrose increased the FB2 production of A. niger, whereas addition of glycerol reduced FB2 production. All three water activity lowering solutes reduced the fumonisin production of the Fusarium species. Conclusion The present study shows that the regulation of fumonisin production is very different in A. niger and Fusarium, and that food and feeds preserved by addition of sugar or salts may be good substrates for fumonisin B2 production by A. niger.

  19. Strain selection and medium optimization for glucoamylase production from industrial potato waste by Aspergillus niger.

    Science.gov (United States)

    Izmirlioglu, Gulten; Demirci, Ali

    2016-06-01

    Glucoamylase is one of the most common enzymes used in the food industry to break down starch into its monomers. Glucoamylase production and its activity are highly dependent on medium composition. Starch is well known as a glucoamylase inducer, and utilization of industrial starchy potato waste is an inexpensive way of improving glucoamylase production. Since glucoamylase production is highly dependent on medium composition, in this study medium optimization for glucoamylase production was considered to enhance glucoamylase activity. Among the evaluated microbial species, Aspergillus niger van Tieghem was found to be the best glucoamylase-producing fungus. The Plackett-Burman design was used to screen various medium ingredients, and malt extract, FeSO4 .7H2 O and CaCl2 ·2H2 O were found to have significant effects on glucoamylase production. Finally, malt extract, FeSO4 .7H2 O and CaCl2 .2H2 O were optimized by using a central composite design of response surface methodology. The results showed that the optimal medium composition for A. niger van Tieghem was 50 g L(-1) industrial waste potato mash supplemented with 51.82 g L(-1) malt extract, 9.27 g L(-1) CaCl2 ·2H2 O and 0.50 g L(-1) FeSO4 .7H2 O. At the end of optimization, glucoamylase activity and glucose production were improved 126% and 98% compared to only industrial waste potato mash basal medium; 274.4 U mL(-1) glucoamylase activity and 41.7 g L(-1) glucose levels were achieved, respectively. © 2015 Society of Chemical Industry. © 2015 Society of Chemical Industry.

  20. Biochar Enhances Aspergillus niger Rock Phosphate Solubilization by Increasing Organic Acid Production and Alleviating Fluoride Toxicity

    Science.gov (United States)

    Mendes, Gilberto de Oliveira; Zafra, David Lopez; Vassilev, Nikolay Bojkov; Silva, Ivo Ribeiro; Ribeiro, José Ivo

    2014-01-01

    During fungal rock phosphate (RP) solubilization, a significant quantity of fluoride (F−) is released together with phosphorus (P), strongly inhibiting the process. In the present study, the effect of two F− adsorbents [activated alumina (Al2O3) and biochar] on RP solubilization by Aspergillus niger was examined. Al2O3 adsorbed part of the F− released but also adsorbed soluble P, which makes it inappropriate for microbial RP solubilization systems. In contrast, biochar adsorbed only F− while enhancing phosphate solubilization 3-fold, leading to the accumulation of up to 160 mg of P per liter. By comparing the values of F− measured in solution at the end of incubation and those from a predictive model, it was estimated that up to 19 mg of F− per liter can be removed from solution by biochar when added at 3 g liter−1 to the culture medium. Thus, biochar acted as an F− sink during RP solubilization and led to an F− concentration in solution that was less inhibitory to the process. In the presence of biochar, A. niger produced larger amounts of citric, gluconic, and oxalic acids, whether RP was present or not. Our results show that biochar enhances RP solubilization through two interrelated processes: partial removal of the released F− and increased organic acid production. Given the importance of organic acids for P solubilization and that most of the RPs contain high concentrations of F−, the proposed solubilization system offers an important technological improvement for the microbial production of soluble P fertilizers from RP. PMID:24610849

  1. Structural features of sugars that trigger or support conidial germination in the filamentous fungus Aspergillus niger.

    Science.gov (United States)

    Hayer, Kimran; Stratford, Malcolm; Archer, David B

    2013-11-01

    The asexual spores (conidia) of Aspergillus niger germinate to produce hyphae under appropriate conditions. Germination is initiated by conidial swelling and mobilization of internal carbon and energy stores, followed by polarization and emergence of a hyphal germ tube. The effects of different pyranose sugars, all analogues of d-glucose, on the germination of A. niger conidia were explored, and we define germination as the transition from a dormant conidium into a germling. Within germination, we distinguish two distinct stages, the initial swelling of the conidium and subsequent polarized growth. The stage of conidial swelling requires a germination trigger, which we define as a compound that is sensed by the conidium and which leads to catabolism of d-trehalose and isotropic growth. Sugars that triggered germination and outgrowth included d-glucose, d-mannose, and d-xylose. Sugars that triggered germination but did not support subsequent outgrowth included d-tagatose, d-lyxose, and 2-deoxy-d-glucose. Nontriggering sugars included d-galactose, l-glucose, and d-arabinose. Certain nontriggering sugars, including d-galactose, supported outgrowth if added in the presence of a complementary triggering sugar. This division of functions indicates that sugars are involved in two separate events in germination, triggering and subsequent outgrowth, and the structural features of sugars that support each, both, or none of these events are discussed. We also present data on the uptake of sugars during the germination process and discuss possible mechanisms of triggering in the absence of apparent sugar uptake during the initial swelling of conidia.

  2. Biological Control of Meloidogyne incognita by Aspergillus niger F22 Producing Oxalic Acid.

    Science.gov (United States)

    Jang, Ja Yeong; Choi, Yong Ho; Shin, Teak Soo; Kim, Tae Hoon; Shin, Kee-Sun; Park, Hae Woong; Kim, Young Ho; Kim, Hun; Choi, Gyung Ja; Jang, Kyoung Soo; Cha, Byeongjin; Kim, In Seon; Myung, Eul Jae; Kim, Jin-Cheol

    2016-01-01

    Restricted usage of chemical nematicides has led to development of environmentally safe alternatives. A culture filtrate of Aspergillus niger F22 was highly active against Meloidogyne incognita with marked mortality of second-stage juveniles (J2s) and inhibition of egg hatching. The nematicidal component was identified as oxalic acid by organic acid analysis and gas chromatography-mass spectroscopy (GC-MS). Exposure to 2 mmol/L oxalic acid resulted in 100% juvenile mortality at 1 day after treatment and suppressed egg hatching by 95.6% at 7 days after treatment. Oxalic acid showed similar nematicidal activity against M. hapla, but was not highly toxic to Bursaphelenchus xylophilus. The fungus was incubated on solid medium and dried culture was used for preparation of a wettable powder-type (WP) formulation as an active ingredient. Two WP formulations, F22-WP10 (ai 10%) and oxalic acid-WP8 (ai 8%), were prepared using F22 solid culture and oxalic acid. In a field naturally infested with M. incognita, application of a mixture of F22-WP10 + oxalic acid-WP8 at 1,000- and 500-fold dilutions significantly reduced gall formation on the roots of watermelon plants by 58.8 and 70.7%, respectively, compared to the non-treated control. The disease control efficacy of the mixture of F22-WP10 + oxalic acid-WP8 was significantly higher than that of a chemical nematicide, Sunchungtan (ai 30% fosthiazate). These results suggest that A. niger F22 can be used as a microbial nematicide for the control of root-knot nematode disease.

  3. Biological Control of Meloidogyne incognita by Aspergillus niger F22 Producing Oxalic Acid.

    Directory of Open Access Journals (Sweden)

    Ja Yeong Jang

    Full Text Available Restricted usage of chemical nematicides has led to development of environmentally safe alternatives. A culture filtrate of Aspergillus niger F22 was highly active against Meloidogyne incognita with marked mortality of second-stage juveniles (J2s and inhibition of egg hatching. The nematicidal component was identified as oxalic acid by organic acid analysis and gas chromatography-mass spectroscopy (GC-MS. Exposure to 2 mmol/L oxalic acid resulted in 100% juvenile mortality at 1 day after treatment and suppressed egg hatching by 95.6% at 7 days after treatment. Oxalic acid showed similar nematicidal activity against M. hapla, but was not highly toxic to Bursaphelenchus xylophilus. The fungus was incubated on solid medium and dried culture was used for preparation of a wettable powder-type (WP formulation as an active ingredient. Two WP formulations, F22-WP10 (ai 10% and oxalic acid-WP8 (ai 8%, were prepared using F22 solid culture and oxalic acid. In a field naturally infested with M. incognita, application of a mixture of F22-WP10 + oxalic acid-WP8 at 1,000- and 500-fold dilutions significantly reduced gall formation on the roots of watermelon plants by 58.8 and 70.7%, respectively, compared to the non-treated control. The disease control efficacy of the mixture of F22-WP10 + oxalic acid-WP8 was significantly higher than that of a chemical nematicide, Sunchungtan (ai 30% fosthiazate. These results suggest that A. niger F22 can be used as a microbial nematicide for the control of root-knot nematode disease.

  4. Aspergillus niger β-Glucosidase Has a Cellulase-like Tadpole Molecular Shape

    Science.gov (United States)

    Lima, Marisa A.; Oliveira-Neto, Mario; Kadowaki, Marco Antonio S.; Rosseto, Flavio R.; Prates, Erica T.; Squina, Fabio M.; Leme, Adriana F. P.; Skaf, Munir S.; Polikarpov, Igor

    2013-01-01

    Aspergillus niger is known to secrete large amounts of β-glucosidases, which have a variety of biotechnological and industrial applications. Here, we purified an A. niger β-glucosidase (AnBgl1) and conducted its biochemical and biophysical analyses. Purified enzyme with an apparent molecular mass of 116 kDa forms monomers in solution as judged by native gel electrophoresis and has a pI value of 4.55, as found for most of the fungi of β-glucosidases. Surprisingly, the small angle x-ray experiments reveal that AnBgl1 has a tadpole-like structure, with the N-terminal catalytic domain and C-terminal fibronectin III-like domain (FnIII) connected by the long linker peptide (∼100 amino acid residues) in an extended conformation. This molecular organization resembles the one adopted by other cellulases (such as cellobiohydrolases, for example) that frequently contain a catalytic domain linked to the cellulose-binding module that mediates their binding to insoluble and polymeric cellulose. The reasons why AnBgl1, which acts on the small soluble substrates, has a tadpole molecular shape are not entirely clear. However, our enzyme pulldown assays with different polymeric substrates suggest that AnBgl1 has little or no capacity to bind to and to adsorb cellulose, xylan, and starch, but it has high affinity to lignin. Molecular dynamics simulations suggested that clusters of residues located in the C-terminal FnIII domain interact strongly with lignin fragments. The simulations showed that numerous arginine residues scattered throughout the FnIII surface play an important role in the interaction with lignin by means of cation-π stacking with the lignin aromatic rings. These results indicate that the C-terminal FnIII domain could be operational for immobilization of the enzyme on the cell wall and for the prevention of unproductive binding of cellulase to the biomass lignin. PMID:24064212

  5. Identification of a Novel L-rhamnose Uptake Transporter in the Filamentous Fungus Aspergillus niger

    Science.gov (United States)

    Sloothaak, Jasper; Odoni, Dorett I.; Martins dos Santos, Vitor A. P.; Schaap, Peter J.

    2016-01-01

    The study of plant biomass utilization by fungi is a research field of great interest due to its many implications in ecology, agriculture and biotechnology. Most of the efforts done to increase the understanding of the use of plant cell walls by fungi have been focused on the degradation of cellulose and hemicellulose, and transport and metabolism of their constituent monosaccharides. Pectin is another important constituent of plant cell walls, but has received less attention. In relation to the uptake of pectic building blocks, fungal transporters for the uptake of galacturonic acid recently have been reported in Aspergillus niger and Neurospora crassa. However, not a single L-rhamnose (6-deoxy-L-mannose) transporter has been identified yet in fungi or in other eukaryotic organisms. L-rhamnose is a deoxy-sugar present in plant cell wall pectic polysaccharides (mainly rhamnogalacturonan I and rhamnogalacturonan II), but is also found in diverse plant secondary metabolites (e.g. anthocyanins, flavonoids and triterpenoids), in the green seaweed sulfated polysaccharide ulvan, and in glycan structures from viruses and bacteria. Here, a comparative plasmalemma proteomic analysis was used to identify candidate L-rhamnose transporters in A. niger. Further analysis was focused on protein ID 1119135 (RhtA) (JGI A. niger ATCC 1015 genome database). RhtA was classified as a Family 7 Fucose: H+ Symporter (FHS) within the Major Facilitator Superfamily. Family 7 currently includes exclusively bacterial transporters able to use different sugars. Strong indications for its role in L-rhamnose transport were obtained by functional complementation of the Saccharomyces cerevisiae EBY.VW.4000 strain in growth studies with a range of potential substrates. Biochemical analysis using L-[3H(G)]-rhamnose confirmed that RhtA is a L-rhamnose transporter. The RhtA gene is located in tandem with a hypothetical alpha-L-rhamnosidase gene (rhaB). Transcriptional analysis of rhtA and rha

  6. Comparative Secretome Analysis of Trichoderma reesei and Aspergillus niger during Growth on Sugarcane Biomass

    Science.gov (United States)

    Borin, Gustavo Pagotto; Sanchez, Camila Cristina; de Souza, Amanda Pereira; de Santana, Eliane Silva; de Souza, Aline Tieppo; Leme, Adriana Franco Paes; Squina, Fabio Marcio; Buckeridge, Marcos; Goldman, Gustavo Henrique; Oliveira, Juliana Velasco de Castro

    2015-01-01

    Background Our dependence on fossil fuel sources and concern about the environment has generated a worldwide interest in establishing new sources of fuel and energy. Thus, the use of ethanol as a fuel is advantageous because it is an inexhaustible energy source and has minimal environmental impact. Currently, Brazil is the world's second largest producer of ethanol, which is produced from sugarcane juice fermentation. However, several studies suggest that Brazil could double its production per hectare by using sugarcane bagasse and straw, known as second-generation (2G) bioethanol. Nevertheless, the use of this biomass presents a challenge because the plant cell wall structure, which is composed of complex sugars (cellulose and hemicelluloses), must be broken down into fermentable sugar, such as glucose and xylose. To achieve this goal, several types of hydrolytic enzymes are necessary, and these enzymes represent the majority of the cost associated with 2G bioethanol processing. Reducing the cost of the saccharification process can be achieved via a comprehensive understanding of the hydrolytic mechanisms and enzyme secretion of polysaccharide-hydrolyzing microorganisms. In many natural habitats, several microorganisms degrade lignocellulosic biomass through a set of enzymes that act synergistically. In this study, two fungal species, Aspergillus niger and Trichoderma reesei, were grown on sugarcane biomass with two levels of cell wall complexity, culm in natura and pretreated bagasse. The production of enzymes related to biomass degradation was monitored using secretome analyses after 6, 12 and 24 hours. Concurrently, we analyzed the sugars in the supernatant. Results Analyzing the concentration of monosaccharides in the supernatant, we observed that both species are able to disassemble the polysaccharides of sugarcane cell walls since 6 hours post-inoculation. The sugars from the polysaccharides such as arabinoxylan and β-glucan (that compose the most external

  7. Inhibition of oxidative phosphorylation for enhancing citric acid production by Aspergillus niger.

    Science.gov (United States)

    Wang, Lu; Zhang, Jianhua; Cao, Zhanglei; Wang, Yajun; Gao, Qiang; Zhang, Jian; Wang, Depei

    2015-01-16

    The spore germination rate and growth characteristics were compared between the citric acid high-yield strain Aspergillus niger CGMCC 5751 and A. niger ATCC 1015 in media containing antimycin A or DNP. We inferred that differences in citric acid yield might be due to differences in energy metabolism between these strains. To explore the impact of energy metabolism on citric acid production, the changes in intracellular ATP, NADH and NADH/NAD+ were measured at various fermentation stages. In addition, the effects of antimycin A or DNP on energy metabolism and citric acid production was investigated by CGMCC 5751. By comparing the spore germination rate and the extent of growth on PDA plates containing antimycin A or DNP, CGMCC 5751 was shown to be more sensitive to antimycin A than ATCC 1015. The substrate-level phosphorylation of CGMCC 5751 was greater than that of ATCC 1015 on PDA plates with DNP. DNP at tested concentrations had no apparent effect on the growth of CGMCC 5751. There were no apparent effects on the mycelial morphology, the growth of mycelial pellets or the dry cell mass when 0.2 mg L(-1) antimycin A or 0.1 mg L(-1) DNP was added to medium at the 24-h time point. The concentrations of intracellular ATP, NADH and NADH/NAD+ of CGMCC 5751 were notably lower than those of ATCC 1015 at several fermentation stages. Moreover, at 96 h of fermentation, the citric acid production of CGMCC 5751 reached up to 151.67 g L(-1) and 135.78 g L(-1) by adding 0.2 mg L(-1) antimycin A or 0.1 mg L(-1) DNP, respectively, at the 24-h time point of fermentation. Thus, the citric acid production of CGMCC 5751 was increased by 19.89% and 7.32%, respectively. The concentrations of intracellular ATP, NADH and NADH/NAD+ of the citric acid high-yield strain CGMCC 5751 were notably lower than those of ATCC 1015. The excessive ATP has a strong inhibitory effect on citric acid accumulation by A. niger. Increasing NADH oxidation and appropriately reducing the concentration of

  8. Identification of a Novel L-rhamnose Uptake Transporter in the Filamentous Fungus Aspergillus niger.

    Directory of Open Access Journals (Sweden)

    Jasper Sloothaak

    2016-12-01

    Full Text Available The study of plant biomass utilization by fungi is a research field of great interest due to its many implications in ecology, agriculture and biotechnology. Most of the efforts done to increase the understanding of the use of plant cell walls by fungi have been focused on the degradation of cellulose and hemicellulose, and transport and metabolism of their constituent monosaccharides. Pectin is another important constituent of plant cell walls, but has received less attention. In relation to the uptake of pectic building blocks, fungal transporters for the uptake of galacturonic acid recently have been reported in Aspergillus niger and Neurospora crassa. However, not a single L-rhamnose (6-deoxy-L-mannose transporter has been identified yet in fungi or in other eukaryotic organisms. L-rhamnose is a deoxy-sugar present in plant cell wall pectic polysaccharides (mainly rhamnogalacturonan I and rhamnogalacturonan II, but is also found in diverse plant secondary metabolites (e.g. anthocyanins, flavonoids and triterpenoids, in the green seaweed sulfated polysaccharide ulvan, and in glycan structures from viruses and bacteria. Here, a comparative plasmalemma proteomic analysis was used to identify candidate L-rhamnose transporters in A. niger. Further analysis was focused on protein ID 1119135 (RhtA (JGI A. niger ATCC 1015 genome database. RhtA was classified as a Family 7 Fucose: H+ Symporter (FHS within the Major Facilitator Superfamily. Family 7 currently includes exclusively bacterial transporters able to use different sugars. Strong indications for its role in L-rhamnose transport were obtained by functional complementation of the Saccharomyces cerevisiae EBY.VW.4000 strain in growth studies with a range of potential substrates. Biochemical analysis using L-[3H(G]-rhamnose confirmed that RhtA is a L-rhamnose transporter. The RhtA gene is located in tandem with a hypothetical alpha-L-rhamnosidase gene (rhaB. Transcriptional analysis of rhtA and

  9. Secretion and properties of a hybrid Kluyveromyces lactis-Aspergillus niger β-galactosidase

    Directory of Open Access Journals (Sweden)

    Becerra Manuel

    2006-12-01

    Full Text Available Abstract Background The β-galactosidase from Kluyveromyces lactis is a protein of outstanding biotechnological interest in the food industry and milk whey reutilization. However, due to its intracellular nature, its industrial production is limited by the high cost associated to extraction and downstream processing. The yeast-system is an attractive method for producing many heterologous proteins. The addition of a secretory signal in the recombinant protein is the method of choice to sort it out of the cell, although biotechnological success is not guaranteed. The cell wall acting as a molecular sieve to large molecules, culture conditions and structural determinants present in the protein, all have a decisive role in the overall process. Protein engineering, combining domains of related proteins, is an alternative to take into account when the task is difficult. In this work, we have constructed and analyzed two hybrid proteins from the β-galactosidase of K. lactis, intracellular, and its Aspergillus niger homologue that is extracellular. In both, a heterologous signal peptide for secretion was also included at the N-terminus of the recombinant proteins. One of the hybrid proteins obtained has interesting properties for its biotechnological utilization. Results The highest levels of intracellular and extracellular β-galactosidase were obtained when the segment corresponding to the five domain of K. lactis β-galactosidase was replaced by the corresponding five domain of the A. niger β-galactosidase. Taking into account that this replacement may affect other parameters related to the activity or the stability of the hybrid protein, a thoroughly study was performed. Both pH (6.5 and temperature (40°C for optimum activity differ from values obtained with the native proteins. The stability was higher than the corresponding to the β-galactosidase of K. lactis and, unlike this, the activity of the hybrid protein was increased by the presence

  10. Development of a mutant strain of Aspergillus niger and optimization of some physical factors for improved calcium gluconate production.

    Science.gov (United States)

    Ray, S; Banik, A K

    1994-12-01

    In the course of mutation studies of Aspergillus niger strain AB with ethylene imine (1:4000), a mutant A. niger AB 501, produced greater amount of calcium gluconate in the culture broth (88.0 g/lit) as against the parent strain (36.0 g/lit) by the surface culture method of fermentation. This mutant was then exposed to UV-rays and a mutant, A.niger AB 1801, was found to produce high calcium gluconate in the culture broth (120 g/lit). The optimum cultural conditions for the production of calcium gluconate by A.niger AB 1801 were pH, 6.5; period of incubation, 9 days; volume of medium in 1 litre flask, 150 ml; temperature, 30 degrees C, volume of inoculum, 7.5 ml of cell suspension containing 2.6 x 10(7) spores and age of inoculum, 6 days old spores of A. niger AB 1801. The maximum yield of calcium gluconate to the above conditions was 168 g/lit. The cultural conditions that support maximum cultural growth did not, however, give optimal yield of calcium gluconate because after having yielded the maximum of calcium gluconate the growth of organism continued to increase further.

  11. Synergistic effect of Aspergillus niger and Trichoderma reesei enzyme sets on the saccharification of wheat straw and sugarcane bagasse.

    Science.gov (United States)

    van den Brink, Joost; Maitan-Alfenas, Gabriela Piccolo; Zou, Gen; Wang, Chengshu; Zhou, Zhihua; Guimarães, Valéria Monteze; de Vries, Ronald P

    2014-10-01

    Plant-degrading enzymes can be produced by fungi on abundantly available low-cost plant biomass. However, enzymes sets after growth on complex substrates need to be better understood, especially with emphasis on differences between fungal species and the influence of inhibitory compounds in plant substrates, such as monosaccharides. In this study, Aspergillus niger and Trichoderma reesei were evaluated for the production of enzyme sets after growth on two "second generation" substrates: wheat straw (WS) and sugarcane bagasse (SCB). A. niger and T. reesei produced different sets of (hemi-)cellulolytic enzymes after growth on WS and SCB. This was reflected in an overall strong synergistic effect in releasing sugars during saccharification using A. niger and T. reesei enzyme sets. T. reesei produced less hydrolytic enzymes after growth on non-washed SCB. The sensitivity to non-washed plant substrates was not reduced by using CreA/Cre1 mutants of T. reesei and A. niger with a defective carbon catabolite repression. The importance of removing monosaccharides for producing enzymes was further underlined by the decrease in hydrolytic activities with increased glucose concentrations in WS media. This study showed the importance of removing monosaccharides from the enzyme production media and combining T. reesei and A. niger enzyme sets to improve plant biomass saccharification. Copyright © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  12. Increasing the Performance of Enzymatic Hydrolysis of Rice Straw Using Mixed Crude Cellulases from Trichoderma reesei and Aspergillus niger.

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    Sugeng Winardi

    2010-11-01

    Full Text Available Increasing the Performance of Enzymatic Hydrolysis of Rice Straw Using Mixed Crude Cellulases fromTrichoderma reesei and Aspergillus niger. The objective of this work is to compare the effectiveness of mixed crudeenzyme cellulase from T. reesei and A. niger with commercial enzyme from A. niger, and to investigate effect ofenzyme to substrate ratio to performance of enzymatic hydrolysis of rice straw. The commercial enzyme from FlukaBiochemica was used, and crude enzyme were prepared by solid fermentation with simple media. Before hydrolized,the rice straw was grinded and sieved and then heated at 85 oC with 2% sodium hydroxide for six hours. Hydrolysis wasconducted in 300 mL beaker flask equipped with mechanical stirrer. Samples were analyzed by dinitrosalicylic acidmethod and measured by spectrophotometer. Both of commercial and mixed crude enzyme show that, the higherenzyme to substrate ratio was higher the glucose concentration obtained. However, ratio of glucose obtained to enzymeused become smaller. The mixture of crude enzyme from T. reesesi dan A. niger that produced in this work was twofold more effective to hydrolyze rice straw than using cellulase enzyme of A. niger from Fluka Biochemika

  13. Optimization of productions of cellulolytic enzymes by Aspergillus niger using residue of mango a substrate Otimização da produção de enzimas celulolíticas obtidas de Aspergillus niger utilizando o resíduo da manga como substrato

    National Research Council Canada - National Science Library

    Tamires Carvalho dos Santos; Ingrid Souza Cavalcanti; Renata Cristina Ferreira Bonomo; Nivio Batista Santana; Marcelo Franco

    2011-01-01

    ... (24, 48, 72, 96 and 120 hours) on the kinetic activity of enzymes cellulolytic, produced during the solid state fermentation of waste from the improvement of mango, with the aid of fungus species Aspergillus niger...

  14. The opposite roles of agdA and glaA on citric acid production in Aspergillus niger.

    Science.gov (United States)

    Wang, Lu; Cao, Zhanglei; Hou, Li; Yin, Liuhua; Wang, Dawei; Gao, Qiang; Wu, Zhenqiang; Wang, Depei

    2016-07-01

    Citric acid is produced by an industrial-scale process of fermentation using Aspergillus niger as a microbial cell factory. However, citric acid production was hindered by the non-fermentable isomaltose and insufficient saccharification ability in A. niger when liquefied corn starch was used as a raw material. In this study, A. niger TNA 101ΔagdA was constructed by deletion of the α-glucosidase-encoding agdA gene in A. niger CGMCC 10142 genome using Agrobacterium tumefaciens-mediated transformation. The transformants A. niger OG 1, OG 17, and OG 31 then underwent overexpression of glucoamylase in A. niger TNA 101ΔagdA. The results showed that the α-glucosidase activity of TNA 101ΔagdA was decreased by 62.5 % compared with CGMCC 10142, and isomaltose was almost undetectable in the fermentation broth. The glucoamylase activity of the transformants OG 1 and OG 17 increased by 34.5 and 16.89 % compared with that of TNA 101ΔagdA, respectively. In addition, for the recombinants TNA 101ΔagdA, OG 1 and OG 17, there were no apparent defects in the growth development. Consequently, in comparison with CGMCC 10142, TNA 101ΔagdA and OG 1 decreased the residual reducing sugar by 52.95 and 88.24 %, respectively, and correspondingly increased citric acid production at the end of fermentation by 8.68 and 16.87 %. Citric acid production was further improved by decreasing the non-fermentable residual sugar and increasing utilization rate of corn starch material in A. niger. Besides, the successive saccharification and citric acid fermentation processes were successfully integrated into one step.

  15. Cellulase production by Aspergillus niger in biofilm, solid-state, and submerged fermentations.

    Science.gov (United States)

    Gamarra, Norma N; Villena, Gretty K; Gutiérrez-Correa, Marcel

    2010-06-01

    Cellulase production by Aspergillus niger was compared in three different culture systems: biofilm, solid-state, and submerged fermentation. Biofilm and solid-state fermentations were carried out on perlite as inert support, and lactose was used as a carbon source in the three culture systems. In cryo-scanning electron microscopy, biofilm and solid-state cultures gave similar morphological patterns and confirmed that both spore first attachment and hyphal adhered growth are helped by the production of an adhesive extracellular matrix. Biofilm cultures produced higher cellulase activities than those in submerged and solid-state cultures (1,768, 1,165, and 1,174 U l(-1), respectively). Although biofilm cultures grew less than the other cultures, they produced significantly higher cellulase yields (370, 212, and 217 U g(-1) lactose, respectively) and volumetric productivities (24, 16, and 16 U l(-1) h(-1), respectively). Likewise, endoglucanase and xylanase activities were higher in biofilm cultures. Under the conditions tested, it seems that fungal attached growth on perlite may favor better enzyme production. Biofilms are efficient systems for cellulase production and may replace solid-state fermentation. Biofilm fermentation holds promise for further optimization and development. The results of this work reveal that fungal biofilms may be used for the commercial production of cellulase employing the technology developed for submerged fermentation at high cell densities.

  16. Influence of mechanical stress and surface interaction on the aggregation of Aspergillus niger conidia.

    Science.gov (United States)

    Grimm, L H; Kelly, S; Völkerding, I I; Krull, R; Hempel, D C

    2005-12-30

    Productivity of fungal cultures is closely linked with their morphologic development. Morphogenesis of coagulating filamentous fungi, like Aspergillus niger, starts with aggregation of conidia, also denominated as spores. Several parameters are presumed to control this event, but little is known about their mode of action. Rational process optimization requires models that mirror the underlying reaction mechanisms. An approach in this regard is suggested and supported by experimental data. Aggregation kinetics was examined for the first 15 h of cultivation under different cultivation conditions. Mechanical stress was considered as well as pH-dependent surface interaction. Deliberations were based on a two-step aggregation mechanism. The first aggregation step is only affected by the pH-value, not by the fluid dynamic conditions in the bioreactor. The second aggregation step, in contrast, depends on the pH-value as well as on agitation and aeration induced power input. For the given experimental set-up, agitation had much more influence than aeration. In addition, hyphal growth rate was determined to be the driving force for the second aggregation step. Copyright 2005 Wiley Periodicals, Inc

  17. Enhancement of invertase production by Aspergillus niger OZ-3 using low-intensity static magnetic fields.

    Science.gov (United States)

    Taskin, Mesut; Esim, Nevzat; Genisel, Mucip; Ortucu, Serkan; Hasenekoglu, Ismet; Canli, Ozden; Erdal, Serkan

    2013-01-01

    The aim of this study is to investigate the effect of low-intensity static magnetic fields (SMFs) on invertase activity and growth on different newly identified molds. The most positive effect of SMFs on invertase activity and growth was observed for Aspergillus niger OZ-3. The submerged production of invertase was performed with the spores obtained at the different exposure times (120, 144, 168, and 196 hr) and magnetic field intensities (0.45, 3, 5, 7, and 9 mT). The normal magnetic field of the laboratory was assayed as 0.45 mT (control). Optimization of magnetic field intensity and exposure time significantly increased biomass production and invertase activity compared to 0.45 mT. The maximum invertase activity (51.14 U/mL) and biomass concentration (4.36 g/L) were achieved with the spores obtained at the 144 hr exposure time and 5 mT magnetic field intensity. The effect of low-intensity static magnetic fields (SMFs) on invertase activities of molds was investigated for the first time in the present study. As an additional contribution, a new hyper-invertase-producing mold strain was isolated.

  18. Characterization And Application Of Tannase Produced By Aspergillus Niger ITCC 6514.07 On Pomegranate Rind

    Directory of Open Access Journals (Sweden)

    Anita Srivastava

    2009-12-01

    Full Text Available Extracellular tannase and gallic acid were produced optimally under submerged fermentation at 37 0C, 72 h, pH 5.0, 10 %(v/v inoculum and 4 %(w/v of the agroresidue pomegranate rind (PR powder by an Aspergillus niger isolate. Tannic acid (1 % stimulated the enzyme production by 245.9 % while with 0.5 % glucose, increase was marginal. Tannase production was inhibited by gallic acid and nitrogen sources such as NH4NO3, NH4Cl, KNO3, asparatic acid, urea and EDTA. The partially purified enzyme showed temperature and pH optima of 35 0C and 6.2 respectively which shifted to 40 0C and 5.8 on immobilization in alginate beads. Activity of the enzyme was inhibited by Zn+2, Ca+, Mn+2, Mg+2, Ba+2and Ag+. The immobilized enzyme removed 68.8 % tannin from juice of aonla/myrobalan (Phyllanthus emblica, a tropical fruit, rich in vitamin C and other essential nutrients. The enzymatic treatment of the juice with minimum reduction in vitamin C is encouraging as non enzymatic treatments of myrobalan juice results in vitamin C removal.

  19. [Influence of amaranth on the production of alpha-amylase using Aspergillus niger NRRL 3112].

    Science.gov (United States)

    Mariani, D D; Lorda, G; Balatti, A P

    2000-01-01

    In this paper the influence of the amaranth seed meal and the aeration conditions on the alpha-amylase production by Aspergillus niger NRRL 3112 were studied. The assays of selection of culture medium were carried out in a rotary shaker at 250 rpm and 2.5 cm stroke. The aeration conditions were studied in a mechanically stirred fermentor New Brunswick type. A concentration of alpha-amylase of 2750 U.Dun/ml was achieved at 120 h with a dry weight of 8.0 g/l, using a base medium with 5.0 g/l Amaranthus cruentus seed meal. In the experiment performed in a New Brunswick fermentor, the highest value was 2806 U.Dun/ml. This result was obtained after 120 h, operating at 300 rpm and an airflow of 1 l/l. min. in a limited dissolved oxygen concentration. It was determined that the increase in the agitation rate was not favorable to the enzyme production, despite that an increase was verified in the dissolved oxygen. The morphology of the microorganism, in long and ramified hyphae, was the critical factor to obtain higher levels of alpha-amylase.

  20. Necrotizing mycotic vasculitis with cerebral infarction caused by Aspergillus niger in a horse with acute typholocolitis.

    Science.gov (United States)

    Tunev, S S; Ehrhart, E J; Jensen, H E; Foreman, J H; Richter, R A; Messick, J B

    1999-07-01

    An 18-year-old Morgan mare was presented to the Veterinary Medical Teaching Hospital, University of Illinois, with a 10-day history of watery diarrhea, depression, and dysphagia. On admission, the animal was severely dehydrated, depressed, and unable to swallow and had no clinical signs of diarrhea. The respiratory and heart rate and body temperature were within normal limits. Following fluid therapy, the mare developed severe watery diarrhea and continued to be depressed, incoordinated, and dysphagic. The animal died on the fourth day after admission and was sent to the Laboratories of Veterinary Diagnostic Medicine for necropsy. Gross postmortem findings were consistent with an acute cerebral infarction in the right cerebral hemisphere, an acute necrotizing typhlocolitis, multifocal petechial and ecchymotic hemorrhages, enlarged and congested pars intermedia of the pituitary gland, and marked bilateral adrenocortical hyperplasia with multifocal areas of necrosis and hemorrhage. Histologic evaluation of the affected brain demonstrated an area of coagulative necrosis of the gray matter, with hemorrhage, vasculitis, and thrombosis. There were many fungal hyphae 3.5-6.0 microm, pale basophilic, septate, and occasionally branching at 45 degrees present in the arterial walls and throughout the necrotic tissue. Immunohistochemical analysis revealed Aspergillus niger as the etiologic agent responsible for the mycotic vasculitis and infarction in the brain. Bacteria culture and immunohistochemical staining of the colon and cecum failed to demonstrate specific pathogens.

  1. Biosynthesis, purification and characterization of endoglucanase from a xylanase producing strain Aspergillus niger B03

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    Georgi Todorov Dobrev

    2012-03-01

    Full Text Available An extracellular endoglucanase was isolated from the culture liquid of xylanase producing strain Aspergillus niger B03. The enzyme was purified to a homogenous form, using consecutive ultrafiltration, anion exchange chromatography, and gel filtration. Endoglucanase was a monomer protein with a molecular weight of 26,900 Da determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and 28,800 Da determined by gel filtration. The optimal pH and temperature values for the enzyme action were 3.5 and 65ºC respectively. Endoglucanase was stable at 40ºC, pH 3.0 for 210 min. The substrate specificity of the enzyme was determined with carboxymethyl cellulose, filter paper, and different glycosides. Endoglucanase displayed maximum activity in the case of carboxymethyl cellulose, with a Km value of 21.01 mg/mL. The substrate specificity and the pattern of substrate degradation suggested that the enzyme is an endoglucanase. Endoglucanase showed a synergism with endoxylanase in corn cobs hydrolysis.

  2. Metabolic activity in dormant conidia of Aspergillus niger and developmental changes during conidial outgrowth.

    Science.gov (United States)

    Novodvorska, Michaela; Stratford, Malcolm; Blythe, Martin J; Wilson, Raymond; Beniston, Richard G; Archer, David B

    2016-09-01

    The early stages of development of Aspergillus niger conidia during outgrowth were explored by combining genome-wide gene expression analysis (RNAseq), proteomics, Warburg manometry and uptake studies. Resting conidia suspended in water were demonstrated for the first time to be metabolically active as low levels of oxygen uptake and the generation of carbon dioxide were detected, suggesting that low-level respiratory metabolism occurs in conidia for maintenance. Upon triggering of spore germination, generation of CO2 increased dramatically. For a short period, which coincided with mobilisation of the intracellular polyol, trehalose, there was no increase in uptake of O2 indicating that trehalose was metabolised by fermentation. Data from genome-wide mRNA profiling showed the presence of transcripts associated with fermentative and respiratory metabolism in resting conidia. Following triggering of conidial outgrowth, there was a clear switch to respiration after 25min, confirmed by cyanide inhibition. No effect of SHAM, salicylhydroxamic acid, on respiration suggests electron flow via cytochrome c oxidase. Glucose entry into spores was not detectable before 1h after triggering germination. The impact of sorbic acid on germination was examined and we showed that it inhibits glucose uptake. O2 uptake was also inhibited, delaying the onset of respiration and extending the period of fermentation. In conclusion, we show that conidia suspended in water are not completely dormant and that conidial outgrowth involves fermentative metabolism that precedes respiration. Copyright © 2016. Published by Elsevier Inc.

  3. Microbial leaching of chromite overburden from Sukinda mines, Orissa, India using Aspergillus niger

    Science.gov (United States)

    Biswas, Supratim; Samanta, Saikat; Dey, Rajib; Mukherjee, Siddhartha; Banerjee, Pataki C.

    2013-08-01

    Leaching of nickel and cobalt from two physical grades (S1, 125-190 μm, coarser and S3, 53-75 μm, finer) of chromite overburden was achieved by treating the overburden (2% pulp density) with 21-d culture filtrate of an Aspergillus niger strain grown in sucrose medium. Metal dissolution increases with ore roasting at 600°C and decreasing particle size due to the alteration of microstructural properties involving the conversion of goethite to hematite and the increase in surface area and porosity as evident from X-ray diffraction (XRD), thermogravimetry-differential thermal analysis (DT-TGA), and field emission scanning electron microscopy (FESEM). About 65% Ni and 59% Co were recovered from the roasted S3 ore employing bioleaching against 26.87% Ni and 31.3% Co using an equivalent amount of synthetic oxalic acid under identical conditions. The results suggest that other fungal metabolites in the culture filtrate played a positive role in the bioleaching process, making it an efficient green approach in Ni and Co recovery from lateritic chromite overburden.

  4. Nutritive value of palm oil sludge fermented with Aspergillus niger after stored in different packing materials

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    T Pasaribu

    2001-12-01

    Full Text Available An experiment has been conducted to determine the effect of type of packaging and time of storage on fermented palm oil sludge. The palm oil sludge was fermented with Aspergillus niger, dried, kept in differentpackaging materials (plastic bags, feed bag, and paper bag and stored under room temperatures for 12 weeks. The experiment was assigned in a split plot design (3x7. The parameters measured were water content, crude protein, soluble nitrogen, true protein, in vitro dry matter (IVDMD and true protein digestibilities (IVTPD, total in vitro digestible protein, and the activities of mannanase and cellulase. Results showed that fermented palm oil sludge stored for 12 weeks increased the water content, decreased the true protein and fiber contents, and also reduced the activity of mannanase and cellulase and in vitro dry matter digestibility but no changes on the crude protein content, protein digestibility, and total digestible protein. It was concluded that fermented palm oil sludge should be kept in feed bag under room temperature if to be stored for 12 weeks.

  5. Purification and characterization of endo-xylanases from Aspergillus Niger. III. An enzyme of PL 365

    Energy Technology Data Exchange (ETDEWEB)

    Fournier, R.A.; Frederick, M.M.; Frederick, J.R.; Reilly, P.J.

    1985-04-01

    An endo-xylanase (1,4-..beta..-D-xylan xylanohydrolase, EC 3.2.1.8) from Aspergillus niger was purified to homogeneity by chromatography with Ultrogel AcA 54, SP-Sephadex C-25 at pH 4.5, DEAE-Sephadex A-25 at pH 5.4, Sephadex G-50, and DEAE-Sephadex A-25 at pH 5.15. The enzyme was active on soluble xylan, on insoluble xylan only after arabinosyl-initiated branch points were removed, and on xylooligosaccharides longer than xylotetraose. There was slight activity on carboxymethyl-cellulose, arabinogalactan, glucomannan, and p-nitrophenyl-..beta..-D- glucopyranoside. The main products of the hydrolysis of soluble and insoluble xylan were oligosaccharides of intermediate length, especially the tri- and pentasaccharides. The isolectric point of the enzyme was 3.65. It had a molecular weight of 2.8 x 10/sup 4/ by SDS-gel electrophoresis, and was high in acidic amino acids but low in those containing sulfur. Highest activity in a 20-min assay at pH 5 was between 40 and 45 degrees C, with an activation energy up to 40 degrees C of 11.1 kJ/mol. The optimum pH for activity was at 5.0. The enzyme was strongly activated by Ca/sup 2 +/. 15 references.

  6. Hydroxylation of 1,8-cineole by Mucor ramannianus and Aspergillus niger.

    Science.gov (United States)

    Ramos, Aline de Souza; Ribeiro, Joyce Benzaquem; Teixeira, Bruna Gomes; Ferreira, José Luiz Pinto; Silva, Jefferson Rocha de A; Ferreira, Alexandre do Amaral; de Souza, Rodrigo Octavio Mendonça Alves; Amaral, Ana Claudia F

    2015-03-01

    The monoterpenoid 1,8-cineole is obtained from the leaves of Eucalyptus globulus and it has important biological activities. It is a cheap natural substrate because it is a by-product of the Eucalyptus cultivation for wood and pulp production. In this study, it was evaluated the potential of three filamentous fungi in the biotransformation of 1,8-cineole. The study was divided in two steps: first, reactions were carried out with 1,8-cineole at 1 g/L for 24 h; afterwards, reactions were carried out with substrate at 5 g/L for 5 days. The substrate was hydroxylated into 2-exo-hydroxy-1,8-cineole and 3-exo-hydroxy-1,8-cineole by fungi Mucor ramannianus and Aspergillus niger with high stereoselectivity. Trichoderma harzianum was also tested but no transformation was detected. M. ramannianus led to higher than 99% of conversion within 24 h with a starting high substrate concentration (1 g/L). When substrate was added at 5 g/L, only M. ramannianus was able to catalyze the reaction, but the conversion level was 21.7% after 5 days. Both products have defined stereochemistry and could be used as chiral synthons. Furthermore, biological activity has been described for 3-exo-hydroxy-1,8-cineol. To the best of our knowledge, this is the first report on the use of M. ramannianus in this reaction.

  7. Hydroxylation of 1,8-cineole by Mucor ramannianusand Aspergillus niger

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    Aline de Souza Ramos

    2015-03-01

    Full Text Available The monoterpenoid 1,8-cineole is obtained from the leaves of Eucalyptus globulus and it has important biological activities. It is a cheap natural substrate because it is a by-product of the Eucalyptuscultivation for wood and pulp production. In this study, it was evaluated the potential of three filamentous fungi in the biotransformation of 1,8-cineole. The study was divided in two steps: first, reactions were carried out with 1,8-cineole at 1 g/L for 24 h; afterwards, reactions were carried out with substrate at 5 g/L for 5 days. The substrate was hydroxylated into 2-exo-hydroxy-1,8-cineole and 3-exo-hydroxy-1,8-cineole by fungi Mucor ramannianus and Aspergillus niger with high stereoselectivity. Trichoderma harzianum was also tested but no transformation was detected. M. ramannianus led to higher than 99% of conversion within 24 h with a starting high substrate concentration (1 g/L. When substrate was added at 5 g/L, only M. ramannianuswas able to catalyze the reaction, but the conversion level was 21.7% after 5 days. Both products have defined stereochemistry and could be used as chiral synthons. Furthermore, biological activity has been described for 3-exo-hydroxy-1,8-cineol. To the best of our knowledge, this is the first report on the use of M. ramannianus in this reaction.

  8. Fluoride-tolerant mutants of Aspergillus niger show enhanced phosphate solubilization capacity.

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    Ubiana de Cássia Silva

    Full Text Available P-solubilizing microorganisms are a promising alternative for a sustainable use of P against a backdrop of depletion of high-grade rock phosphates (RPs. Nevertheless, toxic elements present in RPs, such as fluorine, can negatively affect microbial solubilization. Thus, this study aimed at selecting Aspergillus niger mutants efficient at P solubilization in the presence of fluoride (F-. The mutants were obtained by exposition of conidia to UV light followed by screening in a medium supplemented with Ca3(PO42 and F-. The mutant FS1-555 showed the highest solubilization in the presence of F-, releasing approximately 70% of the P contained in Ca3(PO42, a value 1.7 times higher than that obtained for the wild type (WT. The mutant FS1-331 showed improved ability of solubilizing fluorapatites, increasing the solubilization of Araxá, Catalão, and Patos RPs by 1.7, 1.6, and 2.5 times that of the WT, respectively. These mutants also grew better in the presence of F-, indicating that mutagenesis allowed the acquisition of F- tolerance. Higher production of oxalic acid by FS1-331 correlated with its improved capacity for RP solubilization. This mutant represents a significant improvement and possess a high potential for application in solubilization systems with fluoride-rich phosphate sources.

  9. The Growth of Aspergillus Niger on a Wood Based Material with 4 Types of Wall Finishing

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    Subramaniam Menega

    2016-01-01

    Full Text Available Buildings are a vital component in a human’s daily life. It provides shelter from the environment, weather and animals. Mold growth within the building might be caused by the moisture problems which directly act on it such as water leaks or indirect factor such as high humidity levels. This growth causes esthetic problems and deterioration of its wall coatings. Spores from the fungi also cause health problems to humans. The fungus species studied in this research is Aspergillus niger. The material is made of wood and its finishing is thick wallpaper, thin wallpaper, acrylic paint and glycerol based paint. ASTMD5590-00 standard was used to evaluate fungal growth and to determine if non antifungal agent was effective in inhibiting the amount of fungal growth on four types of wall finishing used on wooden walls. This research was conducted without using any antifungal agent. Highest percentage of growth of the fungi was found on acrylic paint, followed by glycerol based paint and thin wallpaper. Thick wall paper shows the least growth of fungi. The maximum growth is visible on day 12 which is more than 60% by all the wall finishing.

  10. Xylanase production by a local fungal isolate, Aspergillus niger USM AI 1 via solid state

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    Ibrahim Che Omar

    2005-03-01

    Full Text Available Isolate USM A1 I which was identified to be Aspergillus niger was selected as a potential producer of xylanase via a solid state fermentation system (SSF using palm kernel cake (PKC as substrate. The modification of the physical conditions of the SSF system indicated that the xylanase activity was 23.97 U/g PKC at the moisture ratio of 1:0.75 of PKC: moistening agent with the inoculum size of 1¥104 spores/ml and cultivated at the ambient temperature (28±3ºC. The supplementation of additional carbon and nitrogen sources in the PKC medium could enhance enzyme productivity. The maximum production of xylanase and growth obtained with the supplementation of xylose at 0.75% (w/w were 25.40 U/g and 1.69 mg glucosamine/ g PKC. Moreover, the presence of NaNO3 at 0.075% (w/w as additional nitrogen source further enhanced xylanase production to 33.99 U/g PKC although the growth remained unchanged at about 1.67 mg glucosa- mine/g PKC. The optimized conditions showed an increased xylanase production by 157% compared to before the optimization of the SSF system. The xylanase productivity was 23.12 U/mg glucosamine after optimization and 11.72 U/mg glucosamine before optimization.

  11. Rubrofusarin from Aspergillus niger GTS01-4 and its biological activity

    Science.gov (United States)

    Megawati, Dewi, Rizna Triana; Mulyani, Hanny; Maryani, Faiza; Lotullung, Puspa Dewi N.; Minarti

    2017-01-01

    During the research for bioactive secondary metabolites from microorganisms, the terestrial fungi Aspergillus niger GTS01-4 has been investigated for the evaluation of antimicrobial and cytotoxic activities using brine shrimp (Artemia salina) lethality test and MCF-7 cell line. Further chromatographic separation and purification of myselium extract resulted in the isolation identified as rubrofusarin (1). The structure elucidation of isolated compound was performed using 1D-NMR, and LCMS. Furthermore, the cytotoxicity of rubrofusarin (1) was evaluated and resulted with IC50 of 11.51 µg/mL against MCF-7 and LC50 of 368.11 µg/mL against brine shrimp, respectively. However, rubrofusarin (1) showed moderate activity against E. coli, S. aureus, and B. subtilis compared to standard antibiotic, streptomycin. The average zone of inhibition was ranged from 6 to 8 mm at a concentration of 100 µg/disc. These results suggest that rubrofusarin could be a potential candidate in the field of anticancer drug discovery from terrestrial fungi.

  12. Exploring sequence characteristics related to high-level production of secreted proteins in Aspergillus niger.

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    Bastiaan A van den Berg

    Full Text Available Protein sequence features are explored in relation to the production of over-expressed extracellular proteins by fungi. Knowledge on features influencing protein production and secretion could be employed to improve enzyme production levels in industrial bioprocesses via protein engineering. A large set, over 600 homologous and nearly 2,000 heterologous fungal genes, were overexpressed in Aspergillus niger using a standardized expression cassette and scored for high versus no production. Subsequently, sequence-based machine learning techniques were applied for identifying relevant DNA and protein sequence features. The amino-acid composition of the protein sequence was found to be most predictive and interpretation revealed that, for both homologous and heterologous gene expression, the same features are important: tyrosine and asparagine composition was found to have a positive correlation with high-level production, whereas for unsuccessful production, contributions were found for methionine and lysine composition. The predictor is available online at http://bioinformatics.tudelft.nl/hipsec. Subsequent work aims at validating these findings by protein engineering as a method for increasing expression levels per gene copy.

  13. Purification and Characterization of Tannin Acyl Hydrolase from Aspergillus niger ATCC 16620

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    Abdulhameed Sabu

    2005-01-01

    Full Text Available Tannin acyl hydrolase produced extracellularly by the fungal strain Aspergillus niger ATTC 16620 in solid state fermentation was purified from the cell free culture broth by ammonium sulphate fractionation followed by DEAE–Sephadex A-50 chromatography. SDS-PAGE analysis indicated that the enzyme protein molecular mass was 168 kDa. Enzyme activity was stable up to the temperature of 40 °C and the enzyme activity was optimal at pH=6. Tannase activity was maximal at 0.01 M concentration of the substrate. The addition of metal ions like Zn2+, Mn2+, Cu2+, Ca2+, Mg2+and Fe2+ inhibited the enzyme activity. Only K+ ions enhanced tannase activity, and an activity of 4.31 U/mL was reported here. Enzyme activity was maximal after 15–20 min of incubation time, with an activity of 3.9 U/mL. Km was found to be 1.03 mM and Vmax=4.25 mmol/min. Since the enzyme is active over a wide range of pH and temperature it could find potential use in the food-processing industry.

  14. Citric acid production from Aspergillus niger MT-4 using hydrolysate extract of the insect Locusta migratoria.

    Science.gov (United States)

    Taskin, Mesut; Tasar, Gani Erhan; Incekara, Umit

    2013-06-01

    Citric acid (CA) is the most important organic acid used in the food and other industries. Locusta migratoria is an insect species, which has rich nutritional composition (especially protein) and cultivated in some countries. Therefore, the present study investigated the usability of hydrolysate extract of L. migratoria biomass as substrate for the production of CA from Aspergillus niger MT-4. The insect extract (IE) was found to be rich in ash (34.9 g/100 g), protein (35.6 g/100 g) and mineral contents. Yeast extract was found to be the most favorable substrate for biomass production, whereas the maximum production of CA (41.8 g/L) was achieved in the medium containing IE. Besides, uniform pellets with the smallest size (4 mm) were observed in IE medium. It was thought that rich magnesium (6.78 g/100 g) and manganese (1.14 g/100 g) contents of IE increased the production of CA, resulting in the formation of small uniform pellets. This is the first report on the effect of protein-rich insect biomasses on the production of CA. In this regard, L. migratoria biomass was tested for the first time as a CA-production substrate.

  15. Citric Acid Production by Aspergillus niger Cultivated on Parkia biglobosa Fruit Pulp.

    Science.gov (United States)

    Auta, Helen Shnada; Abidoye, Khadijat Toyin; Tahir, Hauwa; Ibrahim, Aliyu Dabai; Aransiola, Sesan Abiodun

    2014-01-01

    The study was conducted to investigate the potential of Parkia biglobosa fruit pulp as substrate for citric acid production by Aspergillus niger. Reducing sugar was estimated by 3,5-dinitrosalicylic acid and citric acid was estimated spectrophotometrically using pyridine-acetic anhydride methods. The studies revealed that production parameters (pH, inoculum size, substrate concentration, incubation temperature, and fermentation period) had profound effect on the amount of citric acid produced. The maximum yield was obtained at the pH of 2 with citric acid of 1.15 g/L and reducing sugar content of 0.541 mMol(-1), 3% vegetative inoculum size with citric acid yield of 0.53 g/L and reducing sugar content of 8.87 mMol(-1), 2% of the substrate concentration with citric acid yield of 0.83 g/L and reducing sugar content of 9.36 mMol(-1), incubation temperature of 55°C with citric acid yield of 0.62 g/L and reducing sugar content of 8.37 mMol(-1), and fermentation period of 5 days with citric acid yield of 0.61 g/L and reducing sugar content of 3.70 mMol(-1). The results of this study are encouraging and suggest that Parkia biglobosa pulp can be harnessed at low concentration for large scale citric acid production.

  16. Citric acid production by a novel Aspergillus niger isolate: I. Mutagenesis and cost reduction studies.

    Science.gov (United States)

    Lotfy, Walid A; Ghanem, Khaled M; El-Helow, Ehab R

    2007-12-01

    Ultraviolet-irradiation (UV), ethyl methane sulfonate (EMS) and acridine orange (AO) were used to induce citric acid overproduction mutations in Aspergillus niger UMIP 2564. Among 15, eight of the mutant derivatives, were improved with respect to citric acid production from sucrose in batch cultures. Maximum product yield (60.25%) was recorded by W5, a stable UV mutant, with approximately 3.2-fold increase when compared to the parental wild type strain. In terms of the kinetic parameters for batch fermentation processes, the mutation doubled the specific substrate uptake rate and achieved 4.5- and 7.5-fold improvements in citric acid productivity and specific productivity, respectively. For reduction of the fermentation medium cost, corn steep liquor and calcium phosphate pre-treated beet molasses were successfully used as substituents of nitrogen and carbon sources in the growth medium, respectively. These medium substitutions resulted in a W5 citric acid fermentation culture with a product yield of 74.56%.

  17. Identification, characterization, and partial purification of glucoamylase from Aspergillus niger (syn A. ficuum) NRRL 3135.

    Science.gov (United States)

    Vandersall, A S; Cameron, R G; Nairn, C J; Yelenosky, G; Wodzinski, R J

    1995-01-01

    The crude extracellular extract of Aspergillus niger (syn A. ficuum) NRRL 3135 contains glucoamylase (exo-1,4-alpha-D-glucanohydrolase, EC 3.2.1.2). The enzyme, a glycoprotein, was purified 7-fold by ion-exchange chromatography, chromatofocusing, and conconavalin A affinity chromatography. The molecular weight of the enzyme was estimated to be 90 kDa by SDS-PAGE and gel permeation chromatography. The pI of the enzyme was 3.4. The temperature optimum of the enzyme was 60 degrees C and the pH optimum was 5.0. The Vmax values for soluble starch, maltose, maltotriose, maltotretraose, maltopentaose, and isomaltose were 55.2, 11.7, 32.3, 47.8, 59.2, 12.5 nKat glucose/sec, respectively and the Km values for the same substrates were 0.09%, 0.67 mM, 0.76 mM, 0.76 mM, 0.68 mM, and 122.01 mM, respectively.

  18. Expression and characterization of a cutinase (AnCUT2 from Aspergillus niger

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    Ahmed Al-Tammar Khadijah

    2016-01-01

    Full Text Available Cutin hydrolase (EC 3.1.1.74, an extracellular polyesterase found in pollens, bacteria and fungi, is an efficient catalyst that exhibits hydrolytic activity on a variety of water-soluble esters, synthetic fibers, plastics and triglycerides. Thus, cutinase can be used in various applications such as ester synthesis, bio-scouring, food and detergent industries. Ancut2 is one of five genes encoding cutinases present in the Aspergillus niger ATCC 10574 genome. The cDNA of Ancut2 comprising of an open reading frame of 816 bp encoding a protein of 271 amino acid residues, was isolated and expressed in Pichia pastoris. The partially purified recombinant cutinase exhibited a molecular mass of approximately 40 kDa. The enzyme showed highest activity at 40°C with a preference for acidic pH (5.0-6.0. AnCUT2 showed hydrolytic activity towards various p-nitrophenyl esters with preference towards shorter chain esters such as p-nitrophenyl butyrate (C4. Scanning Electron Microscopy demonstrated that AnCUT2 was capable of modifying surfaces of synthetic polycaprolactone and polyethylene terephthalate plastics. The properties of this enzyme suggest that it may be applied in synthetic fiber modification and fruit processing industries.

  19. Microalgae Harvest through Fungal Pelletization—Co-Culture of Chlorella vulgaris and Aspergillus niger

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    Sarman Oktovianus Gultom

    2014-07-01

    Full Text Available Microalgae harvesting is a labor- and energy-intensive process and new approaches to harvesting microalgae need to be developed in order to decrease the costs. In this study; co-cultivatation of filamentous fungus (Aspergillus niger and microalgae (Chlorella vulgaris to form cell pellets was evaluated under different conditions, including organic carbon source (glucose; glycerol; and sodium acetate concentration; initial concentration of fungal spores and microalgal cells and light. Results showed that 2 g/L of glucose with a 1:300 ratio of fungi to microalgae provided the best culturing conditions for the process to reach >90% of cell harvest efficiency. The results also showed that an organic carbon source was required to sustain the growth of fungi and form the cell pellets. The microalgae/fungi co-cultures at mixotrophic conditions obtained much higher total biomass than pure cultures of each individual strains; indicating the symbiotic relationship between two strains. This can benefit the microbial biofuel production in terms of cell harvest and biomass production.

  20. Utilization of water hyacinth cellulose for production of cellobiase-rich preparation by Aspergillus niger 1.

    Science.gov (United States)

    Ismail, A M; Abdel-Naby, M A; Abdel-Fattah, A F

    1995-01-01

    Production of a cellobiase-rich preparation by Aspergillus niger 1 was achieved using water hyacinth cellulose as the sole carbon source in the culture medium. Production of cellobiase, carboxymethylcellulase (CMC-ase) and filter paper (FP)-cellulase was favoured by controlling the pH of the culture medium during fermentation at 5.0. Sodium citrate (0.5%), sodium phytate (0.1%), Tween-80 (0.2%, v/v) and asparagine (0.07%) had stimulating effects on the productivity of cellobiase, CMC-ase and FP-cellulase. Potassium dihydrogen phosphate doubled the yield of CMC-ase but had a slight effect on FP-cellulase and cellobiase. Wheat bran had a pronounced stimulating effect on the production of cellobiase and CMC-ase. The combined effects of these stimulators resulted in an enzyme preparation rich in cellobiase and contained 18.5, 0.29 and 2.21 U/ml of cellobiase, FP-cellulase and CMC-ase, respectively. A high cellobiase/FP-cellulase ratio of 63.8:1 was thus obtained with the fungal enzyme preparation. The cellobiase activity was maximal at pH 5.0 and showed good thermostability.

  1. Purification and characterization of extracellular phytase from Aspergillus niger ATCC 9142.

    Science.gov (United States)

    Casey, Anne; Walsh, Gary

    2003-01-01

    Extracellular phytase produced by Aspergillus niger ATCC 9142 was purified to homogeneity by employing an initial ultrafiltration step, followed by chromatography using ion exchange, gel filtration and chromatofocusing steps. The purified enzyme was an 84 kDa, monomeric protein. It possessed a temperature optimum of 65 degrees C, and a pH optimum of 5.0. Km and Vmax values of 100 microM and 7 nmol/s, respectively, were recorded and these values fall well within the range of those previously reported for microbial phytases. Substrate specificity studies indicated that, while the enzyme could hydrolyse a range of non-phytate-based phosphorylated substrates, its preferred substrate was phytate. Phytase activity was moderately stimulated in the presence of Mg2+, Mn2+, Cu2+, Cd2+, Hg2+, Zn2+ and F- ions. Activity was not significantly affected by Fe2- or Fe3- and was moderately inhibited by Ca2+. The enzyme displayed higher thermostability at 80 degrees C than did two commercial phytase products. Initial characterisation of the purified enzyme suggested that it could be a potential candidate for use as an animal feed supplement.

  2. Production of polygalacturonase by Aspergillus niger BC23 isolated from Irvingia gabonensis (African mango fruit

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    Ogbonnaya Nwokoro

    2016-01-01

    Full Text Available Polygalacturonase was produced from Aspergillus niger BC 23 which was isolated from spoiled Irvingia gabonensis fruit. The influence of carbon substrates on enzyme production showed that the medium containing sucrose produced a maximum enzyme yield of 38.5 U/mg protein after 72 h. Enzyme productivity in this medium was much higher than in the medium that contained only citrus pectin as the sole carbon source. Medium containing yeast extract as a nitrogen source caused the production of specific enzyme activity of 31.2 U/mg protein. Results on the effect of metal ions on enzyme activity showed that Ca2+ gave a percent relative activity of 214% in comparison to the native enzyme activity. The enzyme showed maximum activity in slight acid and neutral pH media with optimal activity at pH 4.0. Temperature activity profile of the enzyme showed best activity at a temperature of 35ºC. Dried fruit peels were tested for their abilities to support enzyme production in a media devoid of citrus pectin. The best enzyme productivity of 102.3 U/mg protein was achieved after 72 h in the medium containing orange peel and this level was much higher than that achieved when pure carbon sources or citrus pectin alone were used for enzyme production.

  3. An acidic pectin lyase from Aspergillus niger with favourable efficiency in fruit juice clarification.

    Science.gov (United States)

    Xu, S X; Qin, X; Liu, B; Zhang, D Q; Zhang, W; Wu, K; Zhang, Y H

    2015-02-01

    The pectin lyase gene pnl-zj5a from Aspergillus niger ZJ5 was identified and expressed in Pichia pastoris. PNL-ZJ5A was purified by ultrafiltration, anion exchange and gel chromatography. The Km and Vmax values determined using citrus pectin were 0.66 mg ml(-1) and 32.6 μmol min(-1) mg(-1) , respectively. PNL-ZJ5A exhibited optimal activity at 43°C and retained activity over 25-50°C. PNL-ZJ5A was optimally active at pH 5 and effective in apple juice clarification. Compared with controls, PNL-ZJ5A increased the fruit juice yield significantly. Furthermore, PNL-ZJ5A reduced the viscosity of apple juice by 38.8% and increased its transmittance by 86.3%. PNL-ZJ5A combined with a commercial pectin esterase resulted in higher juice volume. © 2014 The Society for Applied Microbiology.

  4. Development of stable flocculent Saccharomyces cerevisiae strain for continuous Aspergillus niger beta-galactosidase production.

    Science.gov (United States)

    Oliveira, Carla; Teixeira, José A; Lima, Nelson; Da Silva, Nancy A; Domingues, Lucília

    2007-04-01

    A flocculent Saccharomyces cerevisiae strain was engineered to stably secrete Aspergillus niger beta-galactosidase in a continuous high-cell-density bioreactor. The delta-sequences from the yeast retrotransposon Ty1 were used as target sites for the integration of the beta-galactosidase expression cassette. High-copy-number transformants were successfully obtained using the delta-integration system together with the dominant selection antibiotic, G418. The integration of multiple copies was confirmed by genomic Southern blot analysis. Integrants with the highest beta-galactosidase levels (approximately eight gene copies) had similar beta-galactosidase activities as a recombinant strain carrying the beta-galactosidase expression cassette in a YEp-based vector. The beta-galactosidase expression cassettes integrated into the yeast genome were stably maintained after eight sequential batch cultures in a nonselective medium. In continuous high-cell-density culture under the same operating conditions, the integrant strain was more stable than the plasmid-carrying strain. To our knowledge, this is the first study of multicopy delta-integrant stability in a continuous bioreactor operating at different dilution rates.

  5. Production of Citric Acid from a New Substrate, Undersized Semolina, by Aspergillus niger

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    Emine Alben

    2004-01-01

    Full Text Available The production of citric acid from fermentation medium (mass per volume ratio 0.01 % of undersized semolina by Aspergillus niger was studied by shake culture method. The effects of initial pH (4.5, 5.5 and 6.5, methanol (volume fraction 1.0, 2.0 and 3.0 % and ammonium nitrate (mass per volume ratio 0.01 % on the production of citric acid were investigated. Citric acid concentration, biomass concentration and the amount of total carbohydrates (as glucose were determined during fermentation period. About 0.04, 0.0448 and 0.0506 g/L of citric acid was produced in fermentation medium with initial pH=4.5, 5.5 and 6.5, respectively. Citric acid and biomass concentrations were increased with the addition of methanol and ammonium nitrate. The yield of citric acid based on consumed sugar was about 21.9 % in the presence of 3.0 % methanol and 0.01 % ammonium nitrate in fermentation medium with initial pH=6.5.

  6. Stabilization of dimeric β-glucosidase from Aspergillus niger via glutaraldehyde immobilization under different conditions.

    Science.gov (United States)

    Vazquez-Ortega, Perla Guadalupe; Alcaraz-Fructuoso, Maria Teresa; Rojas-Contreras, Juan A; López-Miranda, Javier; Fernandez-Lafuente, Roberto

    2018-03-01

    The dimeric enzyme β-glucosidase from Aspergillus niger has been immobilized on different amino-agarose beads at pH 5 and 7, exploiting the versatility of glutaraldehyde. The stability of the free enzyme depended on enzyme concentration. Immobilization via ion exchange improved enzyme stability/activity, depending on the immobilization pH. However, the enzyme was desorbed in 75 mM NaCl at pH 7 and some stability/enzyme concentration dependence still existed. of these biocatalysts with glutaraldehyde increased enzyme stability (e.g. at pH 5, after incubation under conditions where the enzyme just ionically exchanged was fully inactivated, the activity of the glutaraldehyde treated enzyme remained unaltered). Immobilization on glutaraldehyde pre-activated supports yielded a higher increase in enzyme activity, but the stabilization was lower. While when measuring the enzyme activity at pH 4 there were no changes after immobilization, all immobilized enzymes were more active than the free enzyme at pH 6 and 7 (2-3 times). The Ki/Km ratio did not significantly decrease in any immobilized biocatalysts, and in some cases it worsened in a significant way (by a 9 fold factor using preactivated supports). The new biocatalysts are significantly more stable and avoid enzyme subunit desorption, being the immobilization pH a key point in their design. Copyright © 2017 Elsevier Inc. All rights reserved.

  7. Fate and role of ammonium ions during fermentation of citric acid by Aspergillus niger.

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    Papagianni, Maria; Wayman, Frank; Mattey, Michael

    2005-11-01

    Stoichiometric modeling of the early stages of the citric acid fermentation process by Aspergillus niger revealed that ammonium ions combine with a carbon-containing metabolite inside the cell, in a ratio 1:1, to form a nitrogen compound which is then excreted by the mycelium. High-performance liquid chromatography analysis identified glucosamine as the product of the relationship between glucose and ammonium during the early stages of the citric acid fermentation process. Slightly acidic internal pHs, extremely low ammonium ion concentrations inside the cell, and glucosamine synthesis come into direct contradiction with the earlier theory of the ammonium pool inside the cell, regarded as responsible for inhibition of the enzyme phosphofructokinase. At later fermentation stages, when the mycelium is involved in a process of fragmentation and regrowth, the addition of ammonium sulfate leads to a series of events: the formation and secretion of glucosamine in elevated amounts, the short inhibition of citrate synthesis, growth enhancement, the utilization of glucosamine, and finally, the enhancement of citric acid production rates. Obviously, the enzymatic processes underlining the phenomena need to be reexamined. As a by-product of the citric acid fermentation, glucosamine is reported for the first time here. Suitable process manipulations of the system described in this work could lead to successful glucosamine recovery at the point of its highest yield before degradation by the fungus occurs.

  8. An Experimental Study on Citric Acid Production by Aspergillus niger Using Gelidiella acerosa as a Substrate.

    Science.gov (United States)

    Ramesh, Thangavelu; Kalaiselvam, Murugaiyan

    2011-07-01

    Citric acid (CA) is the most important commercial product which is produced by using various sugar substrates in the terrestrial environment. The present study made an attempt to produce citric acid by the fungal strain Aspergillus niger from red seaweed Gelidiella acerosa is the best alternative to sugar substrate in the marine environment. In this study three types of production media were prepared including control (sucrose) by following standard fermentation conditions. The acid production was indicated by the reduction of pH levels. The control medium gave the highest yield of 80 g/l at pH 1.5 and the medium containing crude seaweed powder and other compositions gave the yield of 30 g/l at pH 3.5 whereas the medium containing crude seaweed and 10% sucrose gave the yield of 50 g/l at pH 3.0. When calculating the benefit cost ratio, crude seaweed powder and 10% sucrose yielded 50 g of citric acid at the lower cost of Rs. 35, whereas the other two media gave the yield of 80 and 30 g respectively with the cost of Rs. 77 and 28. In economic point of view, the medium containing seaweed and 10% sucrose showed more benefit with lower cost.

  9. Improving the extraction conditions of endoglucanase produced by Aspergillus niger under solid-state fermentation

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    R. D. P. B. Pirota

    2013-03-01

    Full Text Available Production of cellulases under solid-state fermentation (SSF is a promising technique that can help to reduce costs. Besides optimizing the production process, it is also important to consider enzyme recovery during the extraction step. Here, an experimental design methodology was used to investigate the effects of the operational parameters solid to liquid ratio (1:3, 1:6 and 1:9, stirring rate (80, 120 and 160 rpm, and temperature (10, 22 and 35 °C on the recovery of endoglucanases produced by Aspergillus niger cultivated under SSF. Statistical analysis revealed that only the solid to liquid ratio had a significant influence on endoglucanase extraction. The highest endoglucanase recovery (35.7 U/g was achieved using 0.2 mol/L acetate buffer at pH 4.8, together with a solid to liquid ratio of 1:9 and an agitation time of 10 minutes. In sequential extraction experiments, it was shown that most of the enzyme was recovered during the first extraction. The procedure adopted increased the efficiency of endoglucanase extraction by 70%, emphasizing the importance of selection of suitable operational conditions during SSF processes.

  10. Biodiesel Production by Aspergillus niger Lipase Immobilized on Barium Ferrite Magnetic Nanoparticles

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    Ahmed I. El-Batal

    2016-05-01

    Full Text Available In this study, Aspergillus niger ADM110 fungi was gamma irradiated to produce lipase enzyme and then immobilized onto magnetic barium ferrite nanoparticles (BFN for biodiesel production. BFN were prepared by the citrate sol-gel auto-combustion method and characterized by transmission electron microscopy (TEM, X-ray diffraction (XRD, Fourier transform infrared (FTIR and scanning electron microscopy with energy dispersive analysis of X-ray (SEM/EDAX analysis. The activities of free and immobilized lipase were measured at various pH and temperature values. The results indicate that BFN–Lipase (5% can be reused in biodiesel production without any treatment with 17% loss of activity after five cycles and 66% loss in activity in the sixth cycle. The optimum reaction conditions for biodiesel production from waste cooking oil (WCO using lipase immobilized onto BFN as a catalyst were 45 °C, 4 h and 400 rpm. Acid values of WCO and fatty acid methyl esters (FAMEs were 1.90 and 0.182 (mg KOH/g oil, respectively. The measured flash point, calorific value and cetane number were 188 °C, 43.1 MJ/Kg and 59.5, respectively. The cloud point (−3 °C, pour point (−9 °C, water content (0.091% and sulfur content (0.050%, were estimated as well.

  11. Niger.

    Science.gov (United States)

    1987-06-01

    Niger is two-thirds Sahara desert and the rest savannah with an area irrigated by the Niger River valley. The 6.2 million people are therefore either nomadic herdsmen or subsistence farmers, coping with a hot, dry climate. There are 5 or more ethnic groups, 2 main languages other than the official French, and most people are Muslim. The growth rate is 3.1%; children make up 45% of the population; infant mortality is 145/1000; life expectancy is 44.5 years. The constitutional government has been suspended by a military regime. A multi-layered structure called "development society" has been instituted. Per capita income is about $265. Niger has uranium, coal, iron, tin and phosphates, and farm products include peanuts, millet, sorghum, beans, cotton, rice and cowpeas. Niger received assistance from France, US, West Germany, Canada, Saudi Arabia, as well as international organizations and military assistance from several countries.

  12. Heterologous expression, purification and characterization of nitrilase from Aspergillus niger K10

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    Felsberg Jürgen

    2011-01-01

    Full Text Available Abstract Background Nitrilases attract increasing attention due to their utility in the mild hydrolysis of nitriles. According to activity and gene screening, filamentous fungi are a rich source of nitrilases distinct in evolution from their widely examined bacterial counterparts. However, fungal nitrilases have been less explored than the bacterial ones. Nitrilases are typically heterogeneous in their quaternary structures, forming short spirals and extended filaments, these features making their structural studies difficult. Results A nitrilase gene was amplified by PCR from the cDNA library of Aspergillus niger K10. The PCR product was ligated into expression vectors pET-30(+ and pRSET B to construct plasmids pOK101 and pOK102, respectively. The recombinant nitrilase (Nit-ANigRec expressed in Escherichia coli BL21-Gold(DE3(pOK101/pTf16 was purified with an about 2-fold increase in specific activity and 35% yield. The apparent subunit size was 42.7 kDa, which is approx. 4 kDa higher than that of the enzyme isolated from the native organism (Nit-ANigWT, indicating post-translational cleavage in the enzyme's native environment. Mass spectrometry analysis showed that a C-terminal peptide (Val327 - Asn356 was present in Nit-ANigRec but missing in Nit-ANigWT and Asp298-Val313 peptide was shortened to Asp298-Arg310 in Nit-ANigWT. The latter enzyme was thus truncated by 46 amino acids. Enzymes Nit-ANigRec and Nit-ANigWT differed in substrate specificity, acid/amide ratio, reaction optima and stability. Refolded recombinant enzyme stored for one month at 4°C was fractionated by gel filtration, and fractions were examined by electron microscopy. The late fractions were further analyzed by analytical centrifugation and dynamic light scattering, and shown to consist of a rather homogeneous protein species composed of 12-16 subunits. This hypothesis was consistent with electron microscopy and our modelling of the multimeric nitrilase, which supports an

  13. Occurrence of fungi and cytotoxicity of the species: Aspergillus ochraceus, Aspergillus niger and Aspergillus flavus isolated from the air of hospital wards.

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    Gniadek, Agnieszka; Krzyściak, Paweł; Twarużek, Magdalena; Macura, Anna B

    2017-03-30

    The basic care requirement for patients with weakened immune systems is to create the environment where the risk of mycosis is reduced to a minimum. Between 2007 and 2013 air samples were collected from various wards of a number of hospitals in Kraków, Poland, by means of the collision method using MAS-100 Iso MH Microbial Air Sampler (Merck Millipore, Germany). The air mycobiota contained several species of fungi, and almost 1/3 of it was made up of the species of the Aspergillus genus. Sixty-one strains of species other than A. fumigatus were selected for the research purposes, namely: 28 strains of A. ochraceus, 22 strains of A. niger and 11 strains of A. flavus species. Selected fungi underwent a cytotoxicity evaluation with the application of the MTT colorimetric assay (3-(4,5-dimethylthiazol-2-yl)-2,5- diphenyltetrazolium bromide). The assay assesses cell viability by means of reducing the yellow tetrazolium salt to insoluble formazan. A semi-quantitative scale for cytotoxicity grading was adopted: low cytotoxic effect (+) with half maximal inhibitory concentration (IC50) for values ranging from 31.251 cm2/ml to 7.813 cm2/ml, medium cytotoxic effect (++) for values ranging from 3.906 cm2/ml to 0.977 cm2/ml and the high one (+++) for values ranging from 0.488 cm2/ml to 0.061 cm2/ml. The absence of cytotoxicity was determined when the IC50 values was at ≥ 50. For 48 samples the analyzed fungi displayed the cytotoxic effect with A. ochraceus in 26 out of 28 cases, with 11 strains displaying the high cytotoxic effect. The lowest cytotoxicity was displayed by fungi of A. niger in 13 out of 22 cases, and the major fungi of A. flavus species were toxic (9 out of 11 cases). A half of the fungi displayed the low cytotoxic effect. On the basis of the comparison of average cytotoxicity levels it was determined that there were significant differences in the levels of cytotoxicity of the analyzed fungi. However, such statement may not provide grounds for a definite

  14. Aspergillus niger PA2: a novel strain for extracellular biotransformation of L-tyrosine into L-DOPA.

    Science.gov (United States)

    Agarwal, Pragati; Pareek, Nidhi; Dubey, Swati; Singh, Jyoti; Singh, R P

    2016-05-01

    L-DOPA (3,4-dihydroxyphenyl-L-alanine), an amino acid derivative is the most widely used drug of choice for the treatment of Parkinson's disease and other neurologic injuries. The present study deals with the elevated biochemical transformation of L-tyrosine to L-DOPA by Aspergillus niger PA2, a potent tyrosinase producer, isolated from decomposed food wastes. This appears to be the first report on A. niger as a notable extracellular tyrosinase producer. The extracellular tyrosinase activity produced remarkably higher levels of L-DOPA, i.e. 2.44 mg mL(-1) when the media was supplemented with 5 mg mL(-1) L-tyrosine. The optimum pH for tyrosinase production was 6.0, with the maximal L-DOPA production at the same pH. The product thus produced was analyzed by thin-layer chromatography, UV spectroscopy, high-performance liquid chromatography and Fourier transform infrared spectroscopy, that had denoted this to be L-DOPA. Kinetic parameters viz. Y p/s, Q s and Q p had further indicated the notable levels of production. Thus, Aspergillus niger PA2 could be a promising resource and may be further exploited for large-scale production of L-DOPA.

  15. Genomic analysis of the secretion stress response in the enzyme-producing cell factory Aspergillus niger

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    Robson Geoffrey D

    2007-06-01

    Full Text Available Abstract Background Filamentous fungi such as Aspergillus niger have a high capacity secretory system and are therefore widely exploited for the industrial production of native and heterologous proteins. However, in most cases the yields of non-fungal proteins are significantly lower than those obtained for fungal proteins. One well-studied bottleneck appears to be the result of mis-folding of heterologous proteins in the ER during early stages of secretion, with related stress responses in the host, including the unfolded protein response (UPR. This study aims at uncovering transcriptional and translational responses occurring in A. niger exposed to secretion stress. Results A genome-wide transcriptional analysis of protein secretion-related stress responses was determined using Affymetrix DNA GeneChips and independent verification for selected genes. Endoplasmic reticulum (ER-associated stress was induced either by chemical treatment of the wild-type cells with dithiothreitol (DTT or tunicamycin, or by expressing a human protein, tissue plasminogen activator (t-PA. All of these treatments triggered the UPR, as shown by the expression levels of several well-known UPR target genes. The predicted proteins encoded by most of the up-regulated genes function as part of the secretory system including chaperones, foldases, glycosylation enzymes, vesicle transport proteins, and ER-associated degradation proteins. Several genes were down-regulated under stress conditions and these included several genes that encode secreted enzymes. Moreover, translational regulation under ER stress was investigated by polysomal fractionation. This analysis confirmed the post-transcriptional control of hacA expression and highlighted that differential translation also occurs during ER stress, in particular for some genes encoding secreted proteins or proteins involved in ribosomal biogenesis and assembly. Conclusion This is first genome-wide analysis of both

  16. Gene identification and functional analysis of methylcitrate synthase in citric acid-producing Aspergillus niger WU-2223L.

    Science.gov (United States)

    Kobayashi, Keiichi; Hattori, Takasumi; Honda, Yuki; Kirimura, Kohtaro

    2013-01-01

    Methylcitrate synthase (EC 2.3.3.5; MCS) is a key enzyme of the methylcitric acid cycle localized in the mitochondria of eukaryotic cells and related to propionic acid metabolism. In this study, cloning of the gene mcsA encoding MCS and heterologous expression of it in Escherichia coli were performed for functional analysis of the MCS of citric acid-producing Aspergillus niger WU-2223L. Only one copy of mcsA (1,495 bp) exists in the A. niger WU-2223L chromosome. It encodes a 51-kDa polypeptide consisting of 465 amino acids containing mitochondrial targeting signal peptides. Purified recombinant MCS showed not only MCS activity (27.6 U/mg) but also citrate synthase (EC 2.3.3.1; CS) activity (26.8 U/mg). For functional analysis of MCS, mcsA disruptant strain DMCS-1, derived from A. niger WU-2223L, was constructed. Although A. niger WU-2223L showed growth on propionate as sole carbon source, DMCS-1 showed no growth. These results suggest that MCS is an essential enzyme in propionic acid metabolism, and that the methylcitric acid cycle operates functionally in A. niger WU-2223L. To determine whether MCS makes a contribution to citric acid production, citric acid production tests on DMCS-1 were performed. The amount of citric acid produced from glucose consumed by DMCS-1 in citric acid production medium over 12 d of cultivation was on the same level to that by WU-2223L. Thus it was found that MCS made no contribution to citric acid production from glucose in A. niger WU-2223L, although MCS showed CS activity.

  17. Comprehensive reconstruction and in silico analysis of Aspergillus niger genome-scale metabolic network model that accounts for 1210 ORFs.

    Science.gov (United States)

    Lu, Hongzhong; Cao, Weiqiang; Ouyang, Liming; Xia, Jianye; Huang, Mingzhi; Chu, Ju; Zhuang, Yingping; Zhang, Siliang; Noorman, Henk

    2017-03-01

    Aspergillus niger is one of the most important cell factories for industrial enzymes and organic acids production. A comprehensive genome-scale metabolic network model (GSMM) with high quality is crucial for efficient strain improvement and process optimization. The lack of accurate reaction equations and gene-protein-reaction associations (GPRs) in the current best model of A. niger named GSMM iMA871, however, limits its application scope. To overcome these limitations, we updated the A. niger GSMM by combining the latest genome annotation and literature mining technology. Compared with iMA871, the number of reactions in iHL1210 was increased from 1,380 to 1,764, and the number of unique ORFs from 871 to 1,210. With the aid of our transcriptomics analysis, the existence of 63% ORFs and 68% reactions in iHL1210 can be verified when glucose was used as the only carbon source. Physiological data from chemostat cultivations, 13 C-labeled and molecular experiments from the published literature were further used to check the performance of iHL1210. The average correlation coefficients between the predicted fluxes and estimated fluxes from 13 C-labeling data were sufficiently high (above 0.89) and the prediction of cell growth on most of the reported carbon and nitrogen sources was consistent. Using the updated genome-scale model, we evaluated gene essentiality on synthetic and yeast extract medium, as well as the effects of NADPH supply on glucoamylase production in A. niger. In summary, the new A. niger GSMM iHL1210 contains significant improvements with respect to the metabolic coverage and prediction performance, which paves the way for systematic metabolic engineering of A. niger. Biotechnol. Bioeng. 2017;114: 685-695. © 2016 Wiley Periodicals, Inc. © 2016 Wiley Periodicals, Inc.

  18. Structural features of Aspergillus niger β-galactosidase define its activity against glycoside linkages.

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    Rico-Díaz, Agustín; Ramírez-Escudero, Mercedes; Vizoso-Vázquez, Ángel; Cerdán, M Esperanza; Becerra, Manuel; Sanz-Aparicio, Julia

    2017-06-01

    β-Galactosidases are biotechnologically interesting enzymes that catalyze the hydrolysis or transgalactosylation of β-galactosides. Among them, the Aspergillus niger β-galactosidase (AnβGal) belongs to the glycoside hydrolase family 35 (GH35) and is widely used in the industry due to its high hydrolytic activity which is used to degrade lactose. We present here its three-dimensional structure in complex with different oligosaccharides, to illustrate the structural determinants of the broad specificity of the enzyme against different glycoside linkages. Remarkably, the residues Phe264, Tyr304, and Trp806 make a dynamic hydrophobic platform that accommodates the sugar at subsite +1 suggesting a main role on the recognition of structurally different substrates. Moreover, complexes with the trisaccharides show two potential subsites +2 depending on the substrate type. This feature and the peculiar shape of its wide cavity suggest that AnβGal might accommodate branched substrates from the complex net of polysaccharides composing the plant material in its natural environment. Relevant residues were selected and mutagenesis analyses were performed to evaluate their role in the catalytic performance and the hydrolase/transferase ratio of AnβGal. Thus, we generated mutants with improved transgalactosylation activity. In particular, the variant Y304F/Y355H/N357G/W806F displays a higher level of galacto-oligosaccharides production than the Aspergillus oryzae β-galactosidase, which is the preferred enzyme in the industry owing to its high transferase activity. Our results provide new knowledge on the determinants modulating specificity and the catalytic performance of fungal GH35 β-galactosidases. In turn, this fundamental background gives novel tools for the future improvement of these enzymes, which represent an interesting target for rational design. Structural data are available in PDB database under the accession numbers 5IFP (native form), 5IHR (in complex with 6

  19. Improvement of glucoamylase production for raw-starch digestion in Aspergillus niger F-01 by maltose stearic acid ester.

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    Sun, Haiyan; Peng, Ming

    2017-04-01

    To investigate the role of maltose stearic acid ester (MSAE) for improving raw-starch-digesting glucoamylase (RSDG) production by Aspergillus niger F-01 and its regulatory mechanism. RSDG activity was enhanced 8.3-fold by adding 5 g MSAE to 1 l basal medium at the beginning of the culture. RSDG biosynthesis nearly ceased by adding 100 μg actinomycin D/ml or 50 μg cycloheximide/ml in a 48 h culture in the basal medium. RSDG biosynthesis was restarted by adding MSAE in the culture repressed by actinomycin D. No revival of RSDG synthesis was observed by adding MSAE in the culture repressed by cycloheximide. MSAE improved RSDG production in A. niger by inducing the transcription of RSDG.

  20. Development of a method for efficient cost-effective screening of Aspergillus niger mutants having increased production of glucoamylase.

    Science.gov (United States)

    Zhu, Xudong; Arman, Bessembayev; Chu, Ju; Wang, Yonghong; Zhuang, Yingping

    2017-05-01

    To develop an efficient cost-effective screening process to improve production of glucoamylase in Aspergillus niger. The cultivation of A. niger was achieved with well-dispersed morphology in 48-deep-well microtiter plates, which increased the throughput of the samples compared to traditional flask cultivation. There was a close negative correlation between glucoamylase and its pH of the fermentation broth. A novel high-throughput analysis method using Methyl Orange was developed. When compared to the conventional analysis method using 4-nitrophenyl α-D-glucopyranoside as substrate, a correlation coefficient of 0.96 by statistical analysis was obtained. Using this novel screening method, we acquired a strain with an activity of 2.2 × 10(3) U ml(-1), a 70% higher yield of glucoamylase than its parent strain.

  1. Enhancing saccharification of wheat straw by mixing enzymes from genetically-modified Trichoderma reesei and Aspergillus niger.

    Science.gov (United States)

    Jiang, Yanping; Duarte, Alexandra Vivas; van den Brink, Joost; Wiebenga, Ad; Zou, Gen; Wang, Chengshu; de Vries, Ronald P; Zhou, Zhihua; Benoit, Isabelle

    2016-01-01

    To increase the efficiency of enzymatic hydrolysis for plant biomass conversion into renewable biofuel and chemicals. By overexpressing the point mutation A824 V transcriptional activator Xyr1 in Trichoderma reesei, carboxymethyl cellulase, cellobiosidase and β-D-glucosidase activities of the best mutant were increased from 1.8 IU/ml, 0.1 IU/ml and 0.05 IU/ml to 4.8 IU/ml, 0.4 IU/ml and 0.3 IU/ml, respectively. The sugar yield of wheat straw saccharification by combining enzymes from this mutant and the Aspergillus niger genetically modified strain ΔcreA/xlnR c/araR c was improved up to 7.5 mg/ml, a 229 % increase compared to the combination of wild type strains. Mixing enzymes from T. reesei and A. niger combined with the genetic modification of transcription factors is a promising strategy to increase saccharification efficiency.

  2. Regulation of alternative oxidase at the transcription stage in Aspergillus niger under the conditions of citric acid production.

    Science.gov (United States)

    Hattori, Takasumi; Kino, Kuniki; Kirimura, Kohtaro

    2009-04-01

    The citric acid-producing fungus Aspergillus niger WU-2223L possesses a cyanide-insensitive respiratory pathway catalyzed by alternative oxidase. The regulation of the alternative oxidase under the conditions of citric acid production was determined from the transcription level of the alternative oxidase gene (aox1). PCR and Southern blot analyses revealed that there is only one copy of aox1 on the chromosome of WU-2223L and no homologous gene of aox1. To confirm the regulation stage of alternative oxidase, alternative oxidase activities and aox1 transcription levels were measured under several cultivation conditions, including those for citric acid production. On each cultivation day, the changes in the specific activity of the alternative oxidase were found to be comparable to those in the transcription level of aox1. These results indicate that the activity of the alternative oxidase encoded by aox1 is regulated at the transcription stage under the conditions tested for A. niger WU-2223L.

  3. Activity stabilization of Aspergillus niger and Escherichia coli phytases immobilized on allophanic synthetic compounds and montmorillonite nanoclays.

    Science.gov (United States)

    Menezes-Blackburn, Daniel; Jorquera, Milko; Gianfreda, Liliana; Rao, Maria; Greiner, Ralf; Garrido, Elizabeth; de la Luz Mora, María

    2011-10-01

    The aim of this work was to study the stabilization of the activity of two commercial microbial phytases (Aspergillus niger and Escherichia coli) after immobilization on nanoclays and to establish optimal conditions for their immobilization. Synthetic allophane, synthetic iron-coated allophanes and natural montmorillonite were chosen as solid supports for phytase immobilization. Phytase immobilization patterns at different pH values were strongly dependent on both enzyme and support characteristics. After immobilization, the residual activity of both phytases was higher under acidic conditions. Immobilization of phytases increased their thermal stability and improved resistance to proteolysis, particularly on iron-coated allophane (6% iron oxide), which showed activation energy (E(a)) and activation enthalpy (ΔH(#)) similar to free enzymes. Montmorillonite as well as allophanic synthetic compounds resulted in a good support for immobilization of E. coli phytase, but caused a severe reduction of A. niger phytase activity. Copyright © 2011. Published by Elsevier Ltd.

  4. Protein kinase A signaling and calcium ions are major players in PAF mediated toxicity against Aspergillus niger.

    Science.gov (United States)

    Binder, Ulrike; Benčina, Mojca; Fizil, Ádám; Batta, Gyula; Chhillar, Anil K; Marx, Florentine

    2015-05-08

    The Penicillium chrysogenum antifungal protein PAF is toxic against potentially pathogenic Ascomycetes. We used the highly sensitive aequorin-expressing model Aspergillus niger to identify a defined change in cytoplasmic free Ca(2+) dynamics in response to PAF. This Ca(2+) signature depended on an intact positively charged lysine-rich PAF motif. By combining Ca(2+) measurements in A. niger mutants with deregulated cAMP/protein kinase A (PKA) signaling, we proved the interconnection of Ca(2+) perturbation and cAMP/PKA signaling in the mechanistic function of PAF. A deep understanding of the mode of action of PAF is an invaluable prerequisite for its future application as new antifungal drug. Copyright © 2015 The Authors. Published by Elsevier B.V. All rights reserved.

  5. Purification and characterization of β-Fructosidase with inulinase activity from Aspergillus niger - 245

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    Vinícius D'Arcadia Cruz

    1998-01-01

    Full Text Available Aspergillus niger - 245, a strain isolated from soil samples showed good β-fructosidase activity when inoculated in medium formulated with dahlia extract tubers. The enzyme was purified by precipitation in ammonium sulphate and percolated in DEAE-Sephadex A-50 and CM-cellulose columns, witch showed a single peack in all the purification steps, maintaining the I/S ratio between 0.32 to, 0.39. Optimum pH for inulinase activity (I was between 4.0 - 4.5 and for invertase activity (S between 2.5 and 5.0. The optimum temperature was 60O.C for both activities and no loss in activity was observed when it was maintained at this temperature for 30 min. The Km value was 1.44 and 5.0, respectively, for I and S and Vm value 10.48 and 30.55, respectively. The I activity was strongly inhibited by Hg2+ and Ag+ and 2 x 10-3 M of glucose, but not by fructose at the same concentration. The enzyme showed an exo-action mechanism, acting on the inulin of different origins. In assay conditions total hydrolysis of all the frutans was obtained, although it has shown larger activity on the chicory inulin than that one from artichoke Jerusalem and dahlia, in the first 30 min. The obtained results suggested that the enzyme presented good potential for industrial application in the preparing the fructose syrupsAspergillus niger - 245, isolado do solo mostrou boa atividade de b-frutosidase meio formulado com extrato de tubérculos de dahlia. A enzima foi purificada por precipitação em sulfato de amônia e percolada em colunas de DEAE-Sephadex A-50 e CM-celulose, produzindo um único pico em todas as fases de purificação e mantendo a relação I/S entre 0,32 a 0,39. O pH ótimo para a atividade de inulinase (I foi encontrado entre 4,0 - 4.5 e para a atividade de invertase (S em 2,5 e 5,0. A temperatura ótima foi de 60O.C para ambas as atividades e nenhuma perda foi observada quando mantida nesta temperatura por 30 min. Os valores de Km foram de 1,44 e 5

  6. Adsorption of amyloglucosidase from Aspergillus niger NRRL 3122 using ion exchange resin

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    Ana Paula Manera

    2008-10-01

    Full Text Available Amyloglucosidase enzyme was produced by Aspergillus niger NRRL 3122 from solid-state fermentation, using deffated rice bran as substrate. The effects of process parameters (pH, temperature in the equilibrium partition coefficient for the system amyloglucosidase - resin DEAE-cellulose were investigated, aiming at obtaining the optimum conditions for a subsequent purification process. The highest partition coefficients were obtained using 0.025M Tris-HCl buffer, pH 8.0 and 25ºC. The conditions that supplied the highest partition coefficient were specified, the isotherm that better described the amyloglucosidase process of adsorption obtained. It was observed that the adsorption could be well described by Langmuir equation and the values of Qm and Kd estimated at 133.0 U mL-1 and 15.4 U mL-1, respectively. From the adjustment of the kinetic curves using the fourth-order Runge-Kutta algorithm, the adsorption (k1 and desorption (k2 constants were obtained through optimization by the least square procedure, and the values calculated were 2.4x10-3 mL U-1 min-1 for k1 and 0.037 min-1 for k2 .A enzima amiloglicosidase foi produzida por Aspergillus niger NRRL 3122 através de fermentação em estado sólido, tendo como substrato farelo de arroz desengordurado. Os efeitos dos parâmetros de processo (pH e temperatura no coeficiente de partição no equilíbrio, para o sistema amiloglicosidase - resina DEAE-celulose foram investigados, com o objetivo de se obter as melhores condições para um posterior processo de purificação. Os maiores coeficientes de partição foram obtidos usando tampão Tris-HCl 0,025M pH 8,0 e 25°C. Determinadas as condições que forneceram o maior coeficiente de partição obteve-se a isoterma que melhor descrevia o processo de adsorção de amiloglicosidase. Foi verificado que adsorção pode ser bem descrita pela equação de Langmuir e os valores de Qm e Kd foram estimados em 133,0 U mL-1 e 15,4 U mL-1 respectivamente. A

  7. Novel fungal FAD glucose dehydrogenase derived from Aspergillus niger for glucose enzyme sensor strips.

    Science.gov (United States)

    Sode, Koji; Loew, Noya; Ohnishi, Yosuke; Tsuruta, Hayato; Mori, Kazushige; Kojima, Katsuhiro; Tsugawa, Wakako; LaBelle, Jeffrey T; Klonoff, David C

    2017-01-15

    In this study, a novel fungus FAD dependent glucose dehydrogenase, derived from Aspergillus niger (AnGDH), was characterized. This enzyme's potential for the use as the enzyme for blood glucose monitor enzyme sensor strips was evaluated, especially by investigating the effect of the presence of xylose during glucose measurements. The substrate specificity of AnGDH towards glucose was investigated, and only xylose was found as a competing substrate. The specific catalytic efficiency for xylose compared to glucose was 1.8%. The specific activity of AnGDH for xylose at 5mM concentration compared to glucose was 3.5%. No other sugars were used as substrate by this enzyme. The superior substrate specificity of AnGDH was also demonstrated in the performance of enzyme sensor strips. The impact of spiking xylose in a sample with physiological glucose concentrations on the sensor signals was investigated, and it was found that enzyme sensor strips using AnGDH were not affected at all by 5mM (75mg/dL) xylose. This is the first report of an enzyme sensor strip using a fungus derived FADGDH, which did not show any positive bias at a therapeutic level xylose concentration on the signal for a glucose sample. This clearly indicates the superiority of AnGDH over other conventionally used fungi derived FADGDHs in the application for SMBG sensor strips. The negligible activity of AnGDH towards xylose was also explained on the basis of a 3D structural model, which was compared to the 3D structures of A. flavus derived FADGDH and of two glucose oxidases. Copyright © 2016 Elsevier B.V. All rights reserved.

  8. Control of Acid Phosphatases Expression from Aspergillus niger by Soil Characteristics

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    Ely Nahas

    2015-10-01

    Full Text Available ABSTRACTThis work studied the acid phosphatase (APase activity from culture medium (extracellular, eAPase and mycelial extract (intracellular, iAPase ofAspergillus niger F111. The influence of fungus growth and phosphate concentration of the media on the synthesis and secretion of phosphatase was demonstrated. The effects of pH, substrate concentration and inorganic and organic compounds added to the reaction mixture on APase activity were also studied. Both enzymes were repressed by high concentrations of phosphate. Overexpression of iAPase in relation to eAPase was detected; iAPase activity was 46.1 times higher than eAPase. The maximal activity of eAPase was after 24h of fungus growth and for iAPase was after 96h. Optimal pH and substrate concentrations were 4.5 and 8.0 mM, respectively. Michaelis-Menten constant (Km for the hydrolysis of p-nitrophenyl phosphate was 0.57 mM with Vmax = 14,285.71 U mg-1 mycelium for the iAPase and 0.31 mM with V max = 147.06 U mg-1 mycelium for eAPase. Organic substances had little effect on acid phosphatases when compared with the salts. Both the APases were inhibited by 10 mM KH 2PO4 and 5 mM (NH42MoO4; eAPase was also inhibited by 1 mM CoCl2.

  9. The carbon starvation response of Aspergillus niger during submerged cultivation: Insights from the transcriptome and secretome

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    Nitsche Benjamin M

    2012-08-01

    Full Text Available Abstract Background Filamentous fungi are confronted with changes and limitations of their carbon source during growth in their natural habitats and during industrial applications. To survive life-threatening starvation conditions, carbon from endogenous resources becomes mobilized to fuel maintenance and self-propagation. Key to understand the underlying cellular processes is the system-wide analysis of fungal starvation responses in a temporal and spatial resolution. The knowledge deduced is important for the development of optimized industrial production processes. Results This study describes the physiological, morphological and genome-wide transcriptional changes caused by prolonged carbon starvation during submerged batch cultivation of the filamentous fungus Aspergillus niger. Bioreactor cultivation supported highly reproducible growth conditions and monitoring of physiological parameters. Changes in hyphal growth and morphology were analyzed at distinct cultivation phases using automated image analysis. The Affymetrix GeneChip platform was used to establish genome-wide transcriptional profiles for three selected time points during prolonged carbon starvation. Compared to the exponential growth transcriptome, about 50% (7,292 of all genes displayed differential gene expression during at least one of the starvation time points. Enrichment analysis of Gene Ontology, Pfam domain and KEGG pathway annotations uncovered autophagy and asexual reproduction as major global transcriptional trends. Induced transcription of genes encoding hydrolytic enzymes was accompanied by increased secretion of hydrolases including chitinases, glucanases, proteases and phospholipases as identified by mass spectrometry. Conclusions This study is the first system-wide analysis of the carbon starvation response in a filamentous fungus. Morphological, transcriptomic and secretomic analyses identified key events important for fungal survival and their chronology. The

  10. Isolation of a thermostable acid phytase from Aspergillus niger UFV-1 with strong proteolysis resistance

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    Paulo S. Monteiro

    2015-03-01

    Full Text Available An Aspergillus niger UFV-1 phytase was characterized and made available for industrial application. The enzyme was purified via ultrafiltration followed by acid precipitation, ion exchange and gel filtration chromatography. This protein exhibited a molecular mass of 161 kDa in gel filtration and 81 kDa in sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE, indicating that it may be a dimer. It presented an optimum temperature of 60 °C and optimum pH of 2.0. The KM for sodium phytate hydrolysis was 30.9 mM, while the kcat and kcat/KM were 1.46 ×105 s−1 and 4.7 × 106s−1.M−1, respectively. The purified phytase exhibited broad specificity on a range of phosphorylated compounds, presenting activity on sodium phytate, p-NPP, 2- naphthylphosphate, 1- naphthylphosphate, ATP, phenyl-phosphate, glucose-6-phosphate, calcium phytate and other substrates. Enzymatic activity was slightly inhibited by Mg2+, Cd2+, K+ and Ca2+, and it was drastically inhibited by F−. The enzyme displayed high thermostability, retaining more than 90% activity at 60 °C during 120 h and displayed a t1/2 of 94.5 h and 6.2 h at 70 °C and 80 °C, respectively. The enzyme demonstrated strong resistance toward pepsin and trypsin, and it retained more than 90% residual activity for both enzymes after 1 h treatment. Additionally, the enzyme efficiently hydrolyzed phytate in livestock feed, liberating 15.3 μmol phosphate/mL after 2.5 h of treatment.

  11. Citric Acid Production by Aspergillus niger Cultivated on Parkia biglobosa Fruit Pulp

    Science.gov (United States)

    Abidoye, Khadijat Toyin; Tahir, Hauwa; Ibrahim, Aliyu Dabai; Aransiola, Sesan Abiodun

    2014-01-01

    The study was conducted to investigate the potential of Parkia biglobosa fruit pulp as substrate for citric acid production by Aspergillus niger. Reducing sugar was estimated by 3,5-dinitrosalicylic acid and citric acid was estimated spectrophotometrically using pyridine-acetic anhydride methods. The studies revealed that production parameters (pH, inoculum size, substrate concentration, incubation temperature, and fermentation period) had profound effect on the amount of citric acid produced. The maximum yield was obtained at the pH of 2 with citric acid of 1.15 g/L and reducing sugar content of 0.541 mMol−1, 3% vegetative inoculum size with citric acid yield of 0.53 g/L and reducing sugar content of 8.87 mMol−1, 2% of the substrate concentration with citric acid yield of 0.83 g/L and reducing sugar content of 9.36 mMol−1, incubation temperature of 55°C with citric acid yield of 0.62 g/L and reducing sugar content of 8.37 mMol−1, and fermentation period of 5 days with citric acid yield of 0.61 g/L and reducing sugar content of 3.70 mMol−1. The results of this study are encouraging and suggest that Parkia biglobosa pulp can be harnessed at low concentration for large scale citric acid production. PMID:27433535

  12. The complete biodegradation pathway of ellagitannins by Aspergillus niger in solid-state fermentation.

    Science.gov (United States)

    Ascacio-Valdés, Juan A; Aguilera-Carbó, Antonio F; Buenrostro, José J; Prado-Barragán, Arely; Rodríguez-Herrera, Raúl; Aguilar, Cristóbal N

    2016-04-01

    Our research group has found preliminary evidences of the fungal biodegradation pathway of ellagitannins, revealing first the existence of an enzyme responsible for ellagitannins degradation, which hydrolyzes pomegranate ellagitannins and it was called ellagitannase or elagitannin acyl hydrolase. However, it is necessary to generate new and clear information in order to understand the ellagitannin degradation mechanisms. This work describes the distinctive and unique features of ellagitannin metabolism in fungi. In this study, hydrolysis of pomegranate ellagitannins by Aspergillus niger GH1 was studied by solid-state culture using polyurethane foam as support and pomegranate ellagitannins as substrate. The experiment was performed during 36 h. Results showed that ellagitannin biodegradation started after 6 h of fermentation, reaching the maximal biodegradation value at 18 h. It was observed that ellagitannase activity appeared after 6 h of culture, then, the enzymatic activity was maintained up to 24 h of culture reaching 390.15 U/L, after this period the enzymatic activity decreased. Electrophoretic band for ellagitannase was observed at 18 h. A band obtained using non-denaturing electrophoresis was identified as ellagitannase, then, a tandem analysis to reveal the ellagitannase activity was performed using Petri plate with pomegranate ellagitannins. The extracts were analyzed by HPLC/MS to evaluate ellagitannins degradation. Punicalin, gallagic acid, and ellagic acid were obtained from punicalagin. HPLC/MS analysis identified the gallagic acid as an intermediate molecule and immediate precursor of ellagic acid. The potential application of catabolic metabolism of ellagitannin hydrolysis for ellagic acid production is outlined. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  13. Application of kaolin to improve citric acid production by a thermophilic Aspergillus niger.

    Science.gov (United States)

    Ali, Sikander

    2006-12-01

    Citric acid production by a thermophilic strain of the filamentous fungus Aspergillus niger IIB-6 in a medium containing blackstrap cane molasses was improved by the addition of kaolin to the fermentation medium. The fermentation was run in a 7.5-l stirred bioreactor (60% working volume). The optimal sugar concentration was found to be 150 g/l. Kaolin (1.0 ml) was added to the fermentation medium to enhance volumetric production. The best results in terms of product formation were observed when 15 parts per million (ppm) kaolin was added 24 h after inoculation. With added kaolin, citric acid production was enhanced 2.34-fold, compared to a control fermentation without added kaolin. The length of incubation to attain this product yield was shortened from 168 to 96 h. The comparison of kinetic parameters showed improved citrate synthase activity of the culture (Y (p/x)=7.046 g/g). When the culture grown at various kaolin concentrations was monitored for Q (p), Q (s), and q (p), there was significant improvement in these variables over the control. Specific production by the culture (q (p)=0.073 g/g cells/h) was improved several fold. The addition of kaolin substantially improved the enthalpy (DeltaH (D)=74.5 kJ/mol) and entropy of activation (DeltaS=-174 J/mol/K) for citric acid production, free energies for transition state formation, and substrate binding for sucrose hydrolysis. The performance of fuzzy logic control of the bioreactor was found to be very promising for an improvement ( approximately 4.2-fold) in the production of citric acid (96.88 g/l), which is of value in commercial applications.

  14. “In Silico” Characterization of 3-Phytase A and 3-Phytase B from Aspergillus niger

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    Doris C. Niño-Gómez

    2017-01-01

    Full Text Available Phytases are used for feeding monogastric animals, because they hydrolyze phytic acid generating inorganic phosphate. Aspergillus niger 3-phytase A (PDB: 3K4Q and 3-phytase B (PDB: 1QFX were characterized using bioinformatic tools. Results showed that both enzymes have highly conserved catalytic pockets, supporting their classification as histidine acid phosphatases. 2D structures consist of 43% alpha-helix, 12% beta-sheet, and 45% others and 38% alpha-helix, 12% beta-sheet, and 50% others, respectively, and pI 4.94 and 4.60, aliphatic index 72.25 and 70.26 and average hydrophobicity of −0,304 and −0.330, respectively, suggesting aqueous media interaction. Glycosylation and glycation sites allowed detecting zones that can affect folding and biological activity, suggesting fragmentation. Docking showed that H59 and H63 act as nucleophiles and that D339 and D319 are proton donor residues. MW of 3K4Q (48.84 kDa and 1QFX (50.78 kDa is similar; 1QFX forms homodimers which will originate homotetramers with several catalytic center accessible to the ligand. 3K4Q is less stable (instability index 45.41 than 1QFX (instability index 33.66, but the estimated lifespan for 3K4Q is superior. Van der Waals interactions generate hydrogen bonds between the active center and O2 or H of the phytic acid phosphate groups, providing greater stability to these temporal molecular interactions.

  15. Highly active promoters and native secretion signals for protein production during extremely low growth rates in Aspergillus niger.

    Science.gov (United States)

    Wanka, Franziska; Arentshorst, Mark; Cairns, Timothy C; Jørgensen, Thomas; Ram, Arthur F J; Meyer, Vera

    2016-08-20

    The filamentous ascomycete Aspergillus niger is used in many industrial processes for the production of enzymes and organic acids by batch and fed-batch cultivation. An alternative technique is continuous cultivation, which promises improved yield and optimized pipeline efficiency. In this work, we have used perfusion (retentostat) cultivation to validate two promoters that are suitable for A. niger continuous cultivation of industrially relevant products. Firstly, promoters of genes encoding either an antifungal protein (Panafp) or putative hydrophobin (PhfbD) were confirmed as active throughout retentostat culture by assessing mRNA and protein levels using a luciferase (mluc) reporter system. This demonstrated the anafp promoter mediates a high but temporally variable expression profile, whereas the hfbD promoter mediates a semi-constant, moderate-to-high protein expression during retentostat culture. In order to assess whether these promoters were suitable to produce heterologous proteins during retentostat cultivation, the secreted antifungal protein (AFP) from Aspergillus giganteus, which has many potential biotechnological applications, was expressed in A. niger during retentostat cultivation. Additionally, this assay was used to concomitantly validate that native secretion signals encoded in anafp and hfbD genes can be harnessed for secretion of heterologous proteins. Afp mRNA and protein abundance were comparable to luciferase measurements throughout retentostat cultivation, validating the use of Panafp and PhfbD for perfusion cultivation. Finally, a gene encoding the highly commercially relevant thermal hysteresis protein (THP) was expressed in this system, which did not yield detectable protein. Both hfbD and anafp promoters are suitable for production of useful products in A. niger during perfusion cultivation. These findings provide a platform for further optimisations for high production of heterologous proteins with industrial relevance.

  16. Antifungal activity of lemon, eucalyptus, thyme, oregano, sage and lavender essential oils against Aspergillus niger and Aspergillus tubingensis isolated from grapes

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    Miroslava Císarová

    2016-01-01

    Full Text Available Today, it is very important to find out the protection of products of natural origin as an alternative to synthetic fungicides. The promising alternative is the use of the essential oils (EOs. Essential oils from plants have great potential as a new source of fungicide to control the pathogenic fungi.The main objective of this study was evaluation of the antifungal activity of lemon (Citrus lemon L., eucalyptus (Eucalyptus globulus LABILL., thyme (Thymus vulgaris L., oregano (Origanum vulgare L. sage (Salvia officinalis L. and lavender (Lavandula angustifolia MILLER. EOs against Aspergillus niger and Aspergillus tubingensis isolated from grapes and their ability to affect the growth. It was tested by using the vapor contact with them. At first both tested isolates were identified by using PCR method. Sequence data of 18S rRNA supported the assignment of these isolates to the genus Aspergillus and species A. niger (ITS region: KT824061; RPB2: KT824060 and A. tubingensis (ITS region: KT824062; RPB2: KT824059. Second, EO antifungal activity was evaluated. The effect of the EO volatile phase was confirmed to inhibit growth of A. niger and A tubingensis. EOs were diluted in DMSO (dimethyl sulfoxide final volume of 100 μL. Only 50 μL this solution was distributed on a round sterile filter paper (1 x 1 cm by micropipette, and the paper was placed in the center of the lid of Petri dishes. Dishes were kept in an inverted position. The essential oils with the most significant activity were determined by method of graded concentration of oils - minimum inhibitory doses (MIDs. The most effective tested EOs were oregano and thyme oils, which totally inhibited growth of tested isolates for all days of incubation at 0.625 μL.cm-3 (in air with MFDs 0.125 μL.cm-3 (in air. Lavender EO was less active aginst tested strains (MIDs 0.313 μL.cm-3. The results showed that the tested EOs had antifungal activity, except lemon and eucalyptus. Sage EO was the only

  17. Utilization of Palm Kernel Cake as a Novel Substrate for Phytase Production by Aspergillus niger USM AI1

    OpenAIRE

    Darah Ibrahim; Sheh-Hong Lim

    2014-01-01

    The present study was carried out to optimize the cultural conditions of phytase production by Aspergillus niger USM AI1 using palm kernel cake (PKC) as the substrate in a solid state fermentation (SSF) system. The optimized cultural conditions of 10 g of PKC with a particle size of ≤0.5 mm, an inoculum size of 1x105 spores/ mL, moisture content of 60% (v/w), and a mixing frequency of once every 48 hours produced 4.91±0.17 U/g dried substrate of phytase and 1.14±0.04 mg glucosamine/ g dried s...

  18. EFFECT OF THE INITIAL pH IN THE CELLULASE OF ASPERGILLUS NIGER PRODUCTION IN SOLID-STATE FERMENTATION

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    Gualberto Leon-Revelo

    2017-04-01

    Full Text Available In this work, a kinetic study is presented analyzing the influence of the initial pH on (PFasa cellulase production by an Aspergillus niger strain, cultured in solid-state fermentation. The results showed that the optimum value of initial pH for PFasa production was 4.5. The PFasa activity reached 25.11 UI/gMS in 18 hours and a productivity of 1.40 UI h-1gMS-1 under these initial pH conditions.

  19. Partition of glucose oxidase from Aspergillus niger in aqueous two-phase systems based on salt and polyethylene glycol

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    Jagdish Singh

    2010-10-01

    Full Text Available The aim of this work was to study the isolation of glucose oxidase (GOx from Aspergillus niger in aqueous two phase system consisting of PEG 7500 (150g l-1, potassium phosphate (175 g l-1, K2HPO4 +KH2PO4 and glucose (10 gl-1, the enzyme was partitioned in polymer phase. By sequential extraction GOx (69.2% was recovered in polymer phase by 11.8 fold purification, giving a yield of 129U mg protein-¹.

  20. Citric acid production by Aspergillus niger van. Tieghem MTCC 281 using waste apple pomace as a substrate.

    Science.gov (United States)

    Kumar, Dinesh; Verma, Rachna; Bhalla, T C

    2010-08-01

    A solid state fermentation process was tried for the production of citric acid from apple pomace left after juice extraction using Aspergillus niger van. Tieghem MTCC 281 spores as inoculum (36.8 × 10(4) spores/100 g of pomace). The yield of citric acid was optimized by varying the amount of methanol (1-5% v/w), temperature (25-35°C) and time of incubation (1-7 days) for fermentation process. Optimum yield of citric acid (4.6 g/100 g of pomace) was recorded with 4% (v/w) methanol after 5 days of incubation at 30°C.

  1. Conservation of the active site motif in Aspergillus niger (ficuum) pH 6.0 optimum acid phosphatase and kidney bean purple acid phosphatase.

    Science.gov (United States)

    Mullaney, E J; Ullah, A H

    1998-02-13

    Aspergillus niger (ficuum) and the kidney bean purple acid phosphatases retained all the essential amino acids in the active site despite a low degree of total sequence homology. This high degree of homology in the sequence motif of A. niger fungal acid phosphatase (Apase6) active site with Kidney bean metallo phosphoesterase (KBPAP) and the absence of the RHG-XRXP sequence motif indicates Apase6 to be a metallophosphoesterase rather than a histidine acid phosphatase.

  2. Niger

    DEFF Research Database (Denmark)

    Hahonou, Eric Komlavi

    2015-01-01

    The chapter provides knowledge about the role of non-state actors in security provision in Niger. It argues that it is of upmost importance to dig into the causes of ongoing armed conflicts and volatile situations. It points out the long-term decline of public service provision (including the rol...

  3. An efficient and optimized purification procedure for the superoxide dismutase from Aspergillus niger. Partial characterization of the purified enzyme.

    Science.gov (United States)

    Hatzinikolaou, D G; Tsoukia, C; Kekos, D; Macris, B J

    1997-01-01

    Cu,Zn-superoxide dismutase was isolated from Aspergillus niger mycelia, harvested at the mid-logarithmic growth phase. The purification scheme aimed at the optimization of the ethanol/chloroform extraction (Tsuchihashi extraction) through response surface methodology. Upon optimum extraction conditions, it was possible to obtain electrophoretically pure enzyme preparations, by the application of one step anion exchange chromatography. The enzyme yield of this simple purification procedure was above 75% while the specific activity of the final preparation was among the highest reported for eucariotic microorganisms. The purified enzyme exhibited similar physicochemical characteristics with other Aspergillus sp. superoxide dismutases revealing an apparent tetrameric structure with a subunit molecular weight of 19 kDa, and a pl of 5.95.

  4. Genetic and phenotypic diversity of naturally isolated wild strains of Aspergillus niger with hyper glucose oxidase production

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    MAHMOUD EL-HARIRI

    2015-12-01

    Full Text Available Glucose oxidase (GOx is the basic stone for many of biological industry worldwide. The improvement of GOx production basically depends on selection of hyper producer strain of Aspergillus niger. Selective isolation and screening for natural hyper producer strains of A. niger and sequence analysis of the GOD gene, which is responsible for production of the enzyme, are very effective approaches to investigate the naturally modified strains of A. niger with hyper productive capacity of GOx enzyme. The aims of the current study were selective isolation of naturally hyper GOx producing strains of A. niger and evaluation of their GOx activities under optimized parameters in the laboratory. Five wild Egyptian isolates of A. niger were screened for GOx and catalase activity using two types of modified basal liquid media. The GOx activity was evaluated by high throughout liquid phase system. The isolates showed a variable activity for GOx production ranged from 0 to 28.7 U.ml-1. One isolate coded Strain 7 was negative GOx producer on Vogel's broth medium in comparison to other isolates, while its GOx activity on Cazpek Dox was considered as positive (7.28 U.ml-1. It was concluded that GOx production is affected by three controllable factors – the basal media components, time of incubation, and the strain with its adaption to the media components‎. Also, the catalase activity was tested and it was produced with a different degree of variability, which may be reflected on GOx stability. GOD genes of these wild variant of A. niger were cloned and sequenced to determine intraspecies diversity of GOD between the wild variants. The comparison of isolated wild variants to other reference hyper GOx producer strains of A. niger was performed to determine if the GOD sequence analysis of these strains can be distinguished based on their GOx activity. This is the first report for isolation and detection of naturally A. niger hyper GOx-producer strains with

  5. A biodegradation study of forest biomass by Aspergillus niger F7: correlation between enzymatic activity, hydrolytic percentage and biodegradation index.

    Science.gov (United States)

    Sharma, Nivedita; Kaushal, Richa; Gupta, Rakesh; Kumar, Sanjeev

    2012-04-01

    Aspergillus niger F7 isolated from soil was found to be the potent producer of cellulase and xylanase. The residue of forest species Toona ciliata, Celtris australis, Cedrus deodara and Pinus roxburghii was selected as substrate for biodegradation study due to its easy availability and wide use in industry. It was subjected to alkali (sodium hydroxide) treatment for enhancing its degradation. Biodegradation of forest waste by hydrolytic enzymes (cellulase and xylanase) secreted by A. niger under solid state fermentation (SSF) was explored. SSF of pretreated forest biomass was found to be superior over untreated forest biomass. Highest extracellular enzyme activity of 2201±23.91 U/g by A. niger was shown in pretreated C. australis wood resulting in 6.72±0.20 percent hydrolysis and 6.99±0.23 biodegradation index (BI). The lowest BI of 1.40±0.08 was observed in untreated saw dust of C. deodara having the least enzyme activity of 238±1.36 U/g of dry matter. Biodegradation of forest biomass under SSF was increased many folds when moistening agent i.e. tap water had been replaced with modified basal salt media (BSM). In BSM mediated degradation of forest waste with A. niger, extracellular enzyme activity was increased up to 4089±67.11 U/g of dry matter in turn resulting in higher BI of 15.4±0.41 and percent hydrolysis of 19.38±0.81 in pretreated C. australis wood. A. niger exhibited higher enzyme activity on pretreated biomass when moistened with modified BSM in this study. Statistically a positive correlation has been drawn between these three factors i.e. enzyme activity, BI and percent hydrolysis of forest biomass thus proving their direct relationship with each other.

  6. A biodegradation study of forest biomass by Aspergillus niger F7: correlation between enzymatic activity, hydrolytic percentage and biodegradation index

    Directory of Open Access Journals (Sweden)

    Nivedita Sharma

    2012-06-01

    Full Text Available Aspergillus niger F7 isolated from soil was found to be the potent producer of cellulase and xylanase. The residue of forest species Toona ciliata, Celtris australis, Cedrus deodara and Pinus roxburghii was selected as substrate for biodegradation study due to its easy availability and wide use in industry. It was subjected to alkali (sodium hydroxide treatment for enhancing its degradation. Biodegradation of forest waste by hydrolytic enzymes (cellulase and xylanase secreted by A. niger under solid state fermentation (SSF was explored. SSF of pretreated forest biomass was found to be superior over untreated forest biomass. Highest extracellular enzyme activity of 2201±23.91 U/g by A. niger was shown in pretreated C. australis wood resulting in 6.72±0.20 percent hydrolysis and 6.99±0.23 biodegradation index (BI. The lowest BI of 1.40±0.08 was observed in untreated saw dust of C. deodara having the least enzyme activity of 238±1.36 U/g of dry matter. Biodegradation of forest biomass under SSF was increased many folds when moistening agent i.e. tap water had been replaced with modified basal salt media (BSM. In BSM mediated degradation of forest waste with A. niger, extracellular enzyme activity was increased up to 4089±67.11 U/g of dry matter in turn resulting in higher BI of 15.4±0.41 and percent hydrolysis of 19.38±0.81 in pretreated C. australis wood. A. niger exhibited higher enzyme activity on pretreated biomass when moistened with modified BSM in this study. Statistically a positive correlation has been drawn between these three factors i.e. enzyme activity, BI and percent hydrolysis of forest biomass thus proving their direct relationship with each other.

  7. Proteome analysis of Aspergillus niger: Lactate added in starch-containing medium can increase production of the mycotoxin fumonisin B2 by modifying acetyl-CoA metabolism

    DEFF Research Database (Denmark)

    Sørensen, Louise Marie; Lametsch, Rene; Andersen, Mikael Rørdam

    2009-01-01

    Background Aspergillus niger is a filamentous fungus found in the environment, on foods and feeds and is used as host for production of organic acids, enzymes and proteins. The mycotoxin fumonisin B2 was recently found to be produced by A. niger and hence very little is known about production...... and regulation of this metabolite. Proteome analysis was used with the purpose to reveal how fumonisin B2 production by A. niger is influenced by starch and lactate in the medium. Results Fumonisin B2 production by A. niger was significantly increased when lactate and starch were combined in the medium......). The proteome of A. niger was clearly different during growth on media containing 3% starch, 3% starch + 3% lactate or 3% lactate. The identity of 59 spots was obtained, mainly those showing higher or lower expression levels on medium with starch and lactate. Many of them were enzymes in primary metabolism...

  8. Metabolic changes in transgenic maize mature seeds over-expressing the Aspergillus niger phyA2.

    Science.gov (United States)

    Rao, Jun; Yang, Litao; Guo, Jinchao; Quan, Sheng; Chen, Guihua; Zhao, Xiangxiang; Zhang, Dabing; Shi, Jianxin

    2016-02-01

    Non-targeted metabolomics analysis revealed only intended metabolic changes in transgenic maize over-expressing the Aspergillus niger phyA2. Genetically modified (GM) crops account for a large proportion of modern agriculture worldwide, raising increasingly the public concerns of safety. Generally, according to substantial equivalence principle, if a GM crop is demonstrated to be equivalently safe to its conventional species, it is supposed to be safe. In this study, taking the advantage of an established non-target metabolomic profiling platform based on the combination of UPLC-MS/MS with GC-MS, we compared the mature seed metabolic changes in transgenic maize over-expressing the Aspergillus niger phyA2 with its non-transgenic counterpart and other 14 conventional maize lines. In total, levels of nine out of identified 210 metabolites were significantly changed in transgenic maize as compared with its non-transgenic counterpart, and the number of significantly altered metabolites was reduced to only four when the natural variations were taken into consideration. Notably, those four metabolites were all associated with targeted engineering pathway. Our results indicated that although both intended and non-intended metabolic changes occurred in the mature seeds of this GM maize event, only intended metabolic pathway was found to be out of the range of the natural metabolic variation in the metabolome of the transgenic maize. Therefore, only when natural metabolic variation was taken into account, could non-targeted metabolomics provide reliable objective compositional substantial equivalence analysis on GM crops.

  9. Enhancement of epoxide hydrolase production by (60) Co gamma and UV irradiation mutagenesis of Aspergillus niger ZJB-09103.

    Science.gov (United States)

    Jin, Huo-Xi; OuYang, Xiao-Kun; Hu, Zhong-Ce

    2017-05-01

    An effective epoxide hydrolase (EH) production strain was mutagenized using (60) Co gamma and UV irradiation. Among positive mutant strains, the EH activity of C2-44 reached 33.7 U/g, which was 267% as much as that of the original Aspergillus niger ZJB-09103. Compared with the wild type, there were significant changes in morphology for C2-44, including the color of mycelia on the slants and the shape of conidial head. In addition, glucose and soybean cake were the optimal carbon and nitrogen source in terms of EH activity for the mutant C2-44 instead of soluble starch and peptone for the wild-type strain. The reaction time required to reach 99% enantiomeric excesses of (S)-epichlorohydrin from racemic substrate was shortened significantly by the mutant C2-44. This phenomenon was probably explained by the higher Vmax for hydrolysis of racemic epichlorohydrin by C2-44 compared with Aspergillus niger ZJB-09103. © 2016 International Union of Biochemistry and Molecular Biology, Inc.

  10. Microbial conversion of ginsenoside Rd from Rb1 by the fungus mutant Aspergillus niger strain TH-10a.

    Science.gov (United States)

    Feng, Li; Xu, Chunchun; Li, Zhuo; Li, Jing; Dai, Yulin; Han, Hongxiang; Yu, Shanshan; Liu, Shuying

    2016-05-18

    Ginsenoside Rd, one of the ginsenosides with significant pharmaceutical activities, is getting more and more attractions on its biotransformation. In this study, a novel fungus mutant, the Aspergillus niger strain TH-10a, which can efficiently convert ginsenoside Rd from Rb1, was obtained through screening survival library of LiCl and ultraviolet (UV) irradiation. The transformation product ginsenoside Rd, generated by removing the outer glucose residue from the position C20 of ginsenoside Rb1, was identified through high-performance liquid chromatography (HPLC) analysis. Factors for the microbial culture and biotransformation were investigated in terms of the carbon sources, the nitrogen sources, pH values, and temperatures. This showed that maximum mycelia growth could be obtained at 28°C and pH 6.0 with cellobiose and tryptone as the carbon source and the nitrogen source, respectively. The highest transformation rate (∼86%) has been achieved at 32°C and pH 5.0 with the feeding time of substrate 48 hr. Also, Aspergillus niger strain TH-10a could tolerate even 40 mg/mL ginseng root extract as substrate with 60% bioconversion rate after 72 hr of treatment at the optimal condition. Our results highlight a novel ginsenoside Rd transformation fungus and illuminate its potentially practical application in the pharmaceutical industries.

  11. Effect of temperature and mixing speed on immobilization of crude enzyme from Aspergillus niger on chitosan for hydrolyzing cellulose

    Science.gov (United States)

    Hamzah, Afan; Gek Ela Kumala, P.; Ramadhani, Dwi; Maziyah, Nurul; Rahmah, Laila Nur; Soeprijanto, Widjaja, Arief

    2017-05-01

    Conversion of cellulose into reducing sugar through enzymatic hydrolysis has advantageous because it produces greater product yield, higher selectivity, require less energy, more moderate operating conditions and environment friendly. However, the nature of the enzyme that is difficult to separate and its expensive price become an obstacle. These obstacles can be overcome by immobilizing the enzyme on chitosan material so that the enzyme can be reused. Chitosan is chosen because it is cheap, inert, hydrophilic, and biocompatible. In this research, we use covalent attachment and combination between covalent attachment and cross-linking method for immobilizing crude enzyme. This research was focusing in study of Effect of temperature and mixing speed on Immobilization Enzyme From Aspergillus Niger on Chitosan For Hydrolyzing both soluble (Carboxymethylcellulose) and insoluble Cellulose (coconut husk). This Research was carried out by three main step. First, coconut husk was pre-treated mechanically and chemically, Second, Crude enzyme from Aspergillus niger strain was immobilized on chitosan in various immobilization condition. At last, the pre-treated coconut husk and Carboxymetylcellulose (CMC) were hydrolyzed by immobilized cellulose on chitosan for reducing sugar production. The result revealed that the most reducing sugar produced by immobilized enzyme on chitosan+GDA with immobilization condition at 30 °C and 125 rpm. Enzyme immobilized on chitosan cross-linked with GDA produced more reducing sugar from preteated coconut husk than enzyme immobilized on chitosan.

  12. Optimization and stability of glucoamylase production by recombinant strains of Aspergillus niger in chemostat culture.

    Science.gov (United States)

    Withers, J M; Swift, R J; Wiebe, M G; Robson, G D; Punt, P J; van den Hondel, C A; Trinci, A P

    1998-08-20

    When grown on a medium containing 5 g maltodextrin L-1, Aspergillus niger transformant N402[pAB6-10]B1, which has an additional 20 copies of the glucoamylase (glaA) gene, produced 320 +/- 8 mg (mean +/- S.E.) glucoamylase (GAM) L-1 in batch culture and 373 +/- 9 mg GAM L-1 in maltodextrin-limited chemostat culture at a dilution rate of 0.13 h-1. These values correspond to specific production rates (qp) of 5.6 and 16.0 mg GAM [g biomass]-1 h-1, respectively. In maltodextrin-limited chemostat cultures grown at dilution rates from 0.06 to 0.14 h-1, GAM was produced by B1 in a growth-correlated manner, demonstrating that a continuous flow culture system operated at a high dilution rate is an efficient way of producing this enzyme. In chemostat cultures grown at high dilution rates, GAM production in chemostat cultures was repressed when the limiting nutrient was fructose or xylose, but derepressed when the limiting nutrient was glucose (qp, 12.0), potassium (6.2), ammonium (4.1), phosphate (2.0), magnesium (1.5) or sulphate (0.9). For chemostat cultures grown at a dilution rate of 0.13 h-1, the addition of 5 g mycopeptone L-1 to a glucose-mineral salts medium resulted in a 64% increase in GAM concentration (from 303 +/- 12 to 496 +/- 10 mg GAM L-1) and a 37% increase in specific production rate (from 12.0 +/- 0.4 to 16.4 +/- 1.6 mg GAM [g biomass]-1 h-1). However, although recombinant protein production was stable for at least 948 h (191 generations) when A. niger B1 was grown in chemostat culture on glucose-mineral salts medium, it was stable for less than 136 h (27 generations) on medium containing mycopeptone. The predominant morphological mutants occurring after prolonged chemostat culture were shown to have selective advantage in the chemostat over the parental strain. Compared to their parental strains, two morphological mutants had similar GAM production levels, while a third had a reduced production level. Growth tests and molecular analysis revealed that the

  13. Effect of citrate on Aspergillus niger phytase adsorption and catalytic activity in soil

    Science.gov (United States)

    Mezeli, Malika; Menezes-Blackburn, Daniel; Zhang, Hao; Giles, Courtney; George, Timothy; Shand, Charlie; Lumsdon, David; Cooper, Patricia; Wendler, Renate; Brown, Lawrie; Stutter, Marc; Blackwell, Martin; Darch, Tegan; Wearing, Catherine; Haygarth, Philip

    2015-04-01

    Current developments in cropping systems that promote mobilisation of phytate in agricultural soils, by exploiting plant-root exudation of phytase and organic acids, offer potential for developments in sustainable phosphorus use. However, phytase adsorption to soil particles and phytate complexion has been shown to inhibit phytate dephosphorylation, thereby inhibiting plant P uptake, increasing the risk of this pool contributing to diffuse pollution and reducing the potential benefits of biotechnologies and management strategies aimed to utilise this abundant reserve of 'legacy' phosphorus. Citrate has been seen to increase phytase catalytic efficiency towards complexed forms of phytate, but the mechanisms by which citrate promotes phytase remains poorly understood. In this study, we evaluated phytase (from Aspergillus niger) inactivation, and change in catalytic properties upon addition to soil and the effect citrate had on adsorption of phytase and hydrolysis towards free, precipitated and adsorbed phytate. A Langmuir model was fitted to phytase adsorption isotherms showing a maximum adsorption of 0.23 nKat g-1 (19 mg protein g-1) and affinity constant of 435 nKat gˉ1 (8.5 mg protein g-1 ), demonstrating that phytase from A.niger showed a relatively low affinity for our test soil (Tayport). Phytases were partially inhibited upon adsorption and the specific activity was of 40.44 nKat mgˉ1 protein for the free enzyme and 25.35 nKat mgˉ1 protein when immobilised. The kinetics of adsorption detailed that most of the adsorption occurred within the first 20 min upon addition to soil. Citrate had no effect on the rate or total amount of phytase adsorption or loss of activity, within the studied citrate concentrations (0-4mM). Free phytases in soil solution and phytase immobilised on soil particles showed optimum activity (>80%) at pH 4.5-5.5. Immobilised phytase showed greater loss of activity at pH levels over 5.5 and lower activities at the secondary peak at pH 2

  14. Role of Aspergillus niger in recovery enhancement of valuable metals from produced red mud in Bayer process.

    Science.gov (United States)

    Vakilchap, F; Mousavi, S M; Shojaosadati, S A

    2016-10-01

    Annual worldwide growth rate of red mud (RM) as a hazardous waste has caused serious environmental problems for its disposal in the mining and metallurgy industries. Accordingly, the aim of this study was to investigate biological leaching of RM and recovery of metals using organic acids exerted by Aspergillus niger. Experiments using A. niger were conducted in batch cultures with a pulp density of 2% (w/v) RM under one-step, two-step and spent-medium bioleaching. Based on HPLC results, the major lixiviant was the secretion of organic acids (citric, gluconic, oxalic and malic) by A. niger. Leaching efficiency of metals in the one-step process was the highest and the amounts of leached metals were 69.8%, 60% and 25.4% for Al, Ti and Fe, respectively. The fungal leaching technique demonstrated an adequate recovery of metals, with an efficient and cost-effective means and respect to a reuse of RM for economic and environmental purposes. Copyright © 2016 Elsevier Ltd. All rights reserved.

  15. The antifungal efficacy of nano-metals supported TiO₂ and ozone on the resistant Aspergillus niger spore.

    Science.gov (United States)

    Yu, Kuo-Pin; Huang, Yi-Ting; Yang, Shang-Chun

    2013-10-15

    Recently, antimicrobial efficacy of nano-metals has been extensively investigated. However, most of the related studies focused on the bactericidal effectiveness. Molds, especially their spores, are more resistant than bacteria, and can build a high concentration in houses due to dampness. Therefore, a comprehensive evaluation of the antifungal effectiveness of nano-metals is necessary. In this study, the nano-metals (Ag, Cu and Ni) supported catalysts were successfully prepared by the incipient wetness impregnation method, while the titanium dioxide (Degussa (Evonik) P25) nanoparticle was served as the support. The antifungal experiments of Aspergillus niger spores were conducted on two surfaces (quartz and putty) in the darkness with and without ozone exposure, respectively. The critical Ag concentration to inhibit the germination and growth of A. niger spores of 5 wt% nano Ag catalyst was 65 mg/mL, lower than several cases in previous studies. The inactivation rate constants (k) of A. niger spores on nano-metals supported catalysts in the presence of ozone (k=0.475-0.966 h(-1)) were much higher than those in the absence of ozone (k=0.001-0.268 h(-1)). However, on the surface of TiO₂ particles, no antifungal effect was observed until 6-h exposure to ozone. Consequently, ozone has a synergetic effect on nano-metals antifungal efficacy. Copyright © 2013 Elsevier B.V. All rights reserved.

  16. Improvement on citric acid production in solid-state fermentation by Aspergillus niger LPB BC mutant using citric pulp.

    Science.gov (United States)

    Rodrigues, Cristine; Vandenberghe, Luciana Porto de Souza; Teodoro, Juliana; Pandey, Ashok; Soccol, Carlos Ricardo

    2009-07-01

    Citric acid (CA) production has been conducted through a careful strain selection, physical-chemical optimization and mutation. The aim of this work was to optimize the physical-chemical conditions of CA production by solid-state fermentation (SSF) using the Aspergillus niger LPB BC strain, which was isolated in our laboratory. The parental and mutant strain showed a good production of CA using citric pulp (CP) as a substrate. The physical-chemical parameters were optimized and the best production was reached at 65% moisture, 30 degrees C and pH 5.5. The influence of the addition of commercial and alternative sugars, nitrogen sources, salts, and alcohols was also studied. The best results (445.4 g of CA/kg of CP) were obtained with sugarcane molasses and 4% methanol (v/w). The mutagenesis induction of LPB BC was performed with UV irradiation. Eleven mutant strains were tested in SSF where two mutants showed a higher CA production when compared to the parental strain. A. niger LPB B3 produced 537.6 g of CA/kg of CP on the sixth day of fermentation, while A. niger LPB B6 produced 616.5 g of CA/kg of CP on the fourth day of fermentation, representing a 19.5% and 37% gain, respectively.

  17. Identification and functional analysis of two Golgi-localized UDP-galactofuranose transporters with overlapping functions in Aspergillus niger.

    Science.gov (United States)

    Park, Joohae; Tefsen, Boris; Heemskerk, Marc J; Lagendijk, Ellen L; van den Hondel, Cees A M J J; van Die, Irma; Ram, Arthur F J

    2015-11-02

    Galactofuranose (Galf)-containing glycoconjugates are present in numerous microbes, including filamentous fungi where they are important for morphology, virulence and maintaining cell wall integrity. The incorporation of Galf-residues into galactomannan, galactomannoproteins and glycolipids is carried out by Golgi-localized Galf transferases. The nucleotide sugar donor used by these transferases (UDP-Galf) is produced in the cytoplasm and has to be transported to the lumen of the Golgi by a dedicated nucleotide sugar transporter. Based on homology with recently identified UDP-Galf-transporters in A. fumigatus and A. nidulans, two putative UDP-Galf-transporters in A. niger were found. Their function and localization was determined by gene deletions and GFP-tagging studies, respectively. The two putative UDP-Galf-transporters in A. niger are homologous to each other and are predicted to contain eleven transmembrane domains (UgtA) or ten transmembrane domains (UgtB) due to a reduced length of the C-terminal part of the UgtB protein. The presence of two putative UDP-Galf-transporters in the genome was not unique for A. niger. From the twenty Aspergillus species analysed, nine species contained two additional putative UDP-Galf-transporters. Three of the nine species were outside the Aspergillus section nigri, indication an early duplication of UDP-Galf-transporters and subsequent loss of the UgtB copy in several aspergilli. Deletion analysis of the single and double mutants in A. niger indicated that the two putative UDP-Galf-transporters (named UgtA and UgtB) have a redundant function in UDP-Galf-transport as only the double mutant displayed a Galf-negative phenotype. The Galf-negative phenotype of the double mutant could be complemented by expressing either CFP-UgtA or CFP-UgtB fusion proteins from their endogenous promoters, indicating that both CFP-tagged proteins are functional. Both Ugt proteins co-localize with each other as well as with the GDP

  18. Degradation of pectic polysaccharides in various fruits by pectinase derived from Aspergillus niger

    OpenAIRE

    Kumpoun, Wilawan; MOTOMURA, Yoshie

    2002-01-01

    リンゴやモモの貯蔵病害の一つとして,Aspergillus 菌によるコウジカビ病が知られている。本報では,24種類の果実の果肉から熱水で抽出したペクチン性多糖類に,Aspergillus niger 由来のペクチナーゼを作用させ,その前後の多糖類の分子量分布をゲル濾過によって分析した。得られた溶出パターンの中でピークの変化の程度によって,ペクチン分子のペクチナーゼによる分解の難易を比較した。リンゴ,ウメ,マンゴーではピークの低分子側への変化が大きかったが,アボカド,ドリアン,ブドウ,キーウィー,ビワ,メロン,ライチー,パパイヤ及びランブータンでは変化が小さかった。マンゴーとパパイヤの未熟果と成熟果のペクチン性多糖類のこの酵素による分解性を見たところ,成熟段階に関わらず,マンゴーでは分子量の減少が大きかったが,パパイヤではほとんど変化が見られなかった。ペクチン性多糖類の同一酵素による分解性に差異が生じる原因について,細胞壁多糖の組成や構造の種特異性などの可能性を考察した。...

  19. Aborto por Aspergillus fumigatus e A. niger em bovinos no sul do Brasil

    Directory of Open Access Journals (Sweden)

    Corbellini Luís G.

    2003-01-01

    Full Text Available As infecções micóticas apresentam distribuição mundial e podem causar placentite e aborto em diversas espécies de animais. Entre setembro 2001 e novembro 2002, foram processados no Setor de Patologia Veterinária, Universidade Federal do Rio Grande do Sul, 147 fetos bovinos abortados com o objetivo de avaliar as principais causas de aborto infeccioso bovino no sul do Brasil. Destes, 34 estavam acompanhados da placenta. Aborto micótico foi diagnosticado em cinco casos (3,4% mediante cultivo puro de quatro amostras de Aspergillus fumigatus e uma de A. niger associado a lesões histológicas características de fungo. Os exames virológico, bacteriológico e imunofluorescência direta para Leptospira sp foram negativos nestes casos. A idade dos fetos variava entre 5 e 8 meses de idade. Lesões macroscópicas foram observadas em quatro casos e eram caracterizadas por áreas circulares multifocais branco-acinzentadas na pele, principalmente na região da cabeça e dorso, em dois fetos, lesões nodulares no fígado em um caso e espessamento dos cotilédones em duas placentas enviadas juntamente com os fetos. Lesões histológicas foram observadas principalmente no fígado, pulmão e placenta, caracterizadas por hepatite necrótica multifocal, broncopneumonia supurativa e placentite necrótica. Através da coloração de Grocott hifas septadas foram observadas em duas placentas e nas bordas das lesões necróticas no fígado de um feto. Em dois casos hifas foram observadas somente na placenta e não no feto, salientando-se a importância deste tecido para o diagnóstico de aborto micótico bovino.

  20. Quantification of the fractal nature of mycelial aggregation in Aspergillus niger submerged cultures

    Directory of Open Access Journals (Sweden)

    Papagianni Maria

    2006-02-01

    Full Text Available Abstract Background Fractal geometry estimates have proven useful in studying the growth strategies of fungi in response to different environments on soil or on agar substrates, but their use in mycelia grown submerged is still rare. In the present study, the effects of certain important fermentation parameters, such as the spore inoculum level, phosphate and manganese concentrations in the medium, on mycelial morphology of the citric acid producer Aspergillus niger were determined by fractal geometry. The value of employing fractal geometry to describe mycelial structures was examined in comparison with information from other descriptors including classic morphological parameters derived from image analysis. Results Fractal analysis of distinct morphological forms produced by fermentation conditions that influence fungal morphology and acid production, showed that the two fractal dimensions DBS (box surface dimension and DBM (box mass dimension are very sensitive indexes, capable of describing morphological differences. The two box-counting methods applied (one applied to the whole mass of the mycelial particles and the other applied to their surface only enabled evaluation of fractal dimensions for mycelial particles in this analysis in the region of DBS = 1.20–1.70 and DBM = 1.20–2.70. The global structure of sufficiently branched mycelia was described by a single fractal dimension D, which did not exceed 1.30. Such simple structures are true mass fractals (DBS = DBM = D and they could be young mycelia or dispersed forms of growth produced by very dense spore inocula (108–109 spores/ml or by addition of manganese in the medium. Mycelial clumps and pellets were effectively discriminated by fractal analysis. Fractal dimension values were plotted together with classic morphological parameters derived from image analysis for comparisons. Their sensitivity to treatment was analogous to the sensitivity of classic morphological parameters

  1. Gene encoding a novel invertase from a xerophilic Aspergillus niger strain and production of the enzyme in Pichia pastoris.

    Science.gov (United States)

    Veana, Fabiola; Fuentes-Garibay, José Antonio; Aguilar, Cristóbal Noé; Rodríguez-Herrera, Raúl; Guerrero-Olazarán, Martha; Viader-Salvadó, José María

    2014-09-01

    β-Fructofuranosidases or invertases (EC 3.2.1.26) are enzymes that are widely used in the food industry, where fructose is preferred over sucrose, because it is sweeter and does not crystallize easily. Since Aspergillus niger GH1, an xerophilic fungus from the Mexican semi-desert, has been reported to be an invertase producer, and because of the need for new enzymes with biotechnological applications, in this work, we describe the gene and amino acid sequence of the invertase from A. niger GH1, and the use of a synthetic gene to produce the enzyme in the methylotrophic yeast Pichia pastoris. In addition, the produced invertase was characterized biochemically. The sequence of the invertase gene had a length of 1770 bp without introns, encodes a protein of 589 amino acids, and presented an identity of 93% and 97% with invertases from Aspergillus kawachi IFO 4308 and A. niger B60, respectively. A 4.2 L culture with the constructed recombinant P. pastoris strain showed an extracellular and periplasmic invertase production at 72 h induction of 498 and 3776 invertase units (U), respectively, which corresponds to 1018 U/L of culture medium. The invertase produced had an optimum pH of 5.0, optimum temperature of 60 °C, and specific activity of 3389 U/mg protein, and after storage for 96 h at 4 °C showed 93.7% of its activity. This invertase could be suitable for producing inverted sugar used in the food industry. Copyright © 2014 Elsevier Inc. All rights reserved.

  2. Comprehensive annotation of secondary metabolite biosynthetic genes and gene clusters of Aspergillus nidulans, A. fumigatus, A. niger and A. oryzae

    Science.gov (United States)

    2013-01-01

    Background Secondary metabolite production, a hallmark of filamentous fungi, is an expanding area of research for the Aspergilli. These compounds are potent chemicals, ranging from deadly toxins to therapeutic antibiotics to potential anti-cancer drugs. The genome sequences for multiple Aspergilli have been determined, and provide a wealth of predictive information about secondary metabolite production. Sequence analysis and gene overexpression strategies have enabled the discovery of novel secondary metabolites and the genes involved in their biosynthesis. The Aspergillus Genome Database (AspGD) provides a central repository for gene annotation and protein information for Aspergillus species. These annotations include Gene Ontology (GO) terms, phenotype data, gene names and descriptions and they are crucial for interpreting both small- and large-scale data and for aiding in the design of new experiments that further Aspergillus research. Results We have manually curated Biological Process GO annotations for all genes in AspGD with recorded functions in secondary metabolite production, adding new GO terms that specifically describe each secondary metabolite. We then leveraged these new annotations to predict roles in secondary metabolism for genes lacking experimental characterization. As a starting point for manually annotating Aspergillus secondary metabolite gene clusters, we used antiSMASH (antibiotics and Secondary Metabolite Analysis SHell) and SMURF (Secondary Metabolite Unknown Regions Finder) algorithms to identify potential clusters in A. nidulans, A. fumigatus, A. niger and A. oryzae, which we subsequently refined through manual curation. Conclusions This set of 266 manually curated secondary metabolite gene clusters will facilitate the investigation of novel Aspergillus secondary metabolites. PMID:23617571

  3. Aspergillus niger protein estA defines a new class of fungal esterases within the alfa/beta hydrolase fold superfamily of proteins

    NARCIS (Netherlands)

    Bourne, Y.; Hasper, A.A.; Chahinian, H.; Juin, M.; Graaff, de L.H.

    2004-01-01

    From the fungus Aspergillus niger, we identified a new gene encoding protein EstA, a member of the alpha/beta-hydrolase fold superfamily but of unknown substrate specificity. EstA was overexpressed and its crystal structure was solved by molecular replacement using a lipaseacetylcholinesterase

  4. Aspergillus niger genome-wide analysis reveals a large number of novel alpha-glucan acting enzymes with unexpected expression profiles

    NARCIS (Netherlands)

    Yuan, X.-L.; Kaaij, R.M. van der; Hondel, C.A.M.J.J. van den; Punt, P.J.; Maarel, M.J.E.C. van der; Dijkhuizen, L.; Ram, A.F.J.

    2008-01-01

    The filamentous ascomycete Aspergillus niger is well known for its ability to produce a large variety of enzymes for the degradation of plant polysaccharide material. A major carbon and energy source for this soil fungus is starch, which can be degraded by the concerted action of α-amylase,

  5. The activity of barley alpha-amylase on starch granules is enhanced by fusion of a starch binding domain from Aspergillus niger glucoamylase

    DEFF Research Database (Denmark)

    Juge, N.; Nøhr, J.; Le Gal-Coëffet, M.-F.

    2006-01-01

    High affinity for starch granules of certain amylolytic enzymes is mediated by a separate starch binding domain (SBD). In Aspergillus niger glucoamylase (GA-I), a 70 amino acid O-glycosylated peptide linker connects SBD with the catalytic domain. A gene was constructed to encode barley alpha...

  6. Glucose uptake and growth of glucose-limited chemostat cultures of Aspergillus niger and a disruptant lacking MstA, a high-affinity glucose transporter

    NARCIS (Netherlands)

    Jorgensen, T.R.; vanKuyk, P.A.; Poulsen, B.R.; Ruijter, G.J.G.; Visser, J.; Iversen, J.J.L.

    2007-01-01

    This is a study of high-affinity glucose uptake in Aspergillus niger and the effect of disruption of a high-affinity monosaccharide-transporter gene, mstA. The substrate saturation constant (K-s) of a reference strain was about 15 mu M in glucose-limited chemostat culture. Disruption of mstA

  7. The genome of an industrial workhorse : sequencing of the filamentous fungus Aspergillus niger offers new opportunities for the production of specialty chemicals and enzymes

    Science.gov (United States)

    Dan Cullen

    2007-01-01

    Few microbes compare with the filamentous fungus Aspergillus niger in its ability to produce prodigious amounts of useful chemicals and enzymes. This fungus is the principal source of citric acid for food, beverages and pharmaceuticals and of several important commercial enzymes, including glucoamylase, which is widely used for the conversion of starch to food syrups...

  8. Isolation of a fluffy mutant of Aspergillus niger from chemostat culture and its potential use as a morphologically stable host for protein production

    NARCIS (Netherlands)

    Vondervoort, van de P.J.I.; Poulsen, B.R.; Ruijter, G.J.G.; Schuleit, T.; Visser, J.; Iversen, J.J.L.

    2004-01-01

    Chemostat cultivation of Aspergillus niger and other filamentous fungi is often hindered by the spontaneous appearance of morphologic mutants. Using the Variomixing bioreactor and applying different chemostat conditions we tried to optimize morphologic stability in both ammonium- and glucose-limited

  9. The Aspergillus niger MADS-box transcription factor RlmA is required for cell wall reinforcement in response to cell wall stress

    NARCIS (Netherlands)

    Damveld, R.A.; Arentshorst, M.; Franken, A.; Kuyk, P.A. van; Klis, F.M.; Hondel, C.A.M.J.J. van den; Ram, A.F.J.

    2005-01-01

    In Aspergillus niger, the genes coding for glutamine:fructose-6-phosphate amidotransferase (gfaA) and α-1,3-glucan synthase (agsA) are induced in response to cell wall stress. In silico analysis of the promoter region of the two genes revealed the presence of putative DNA binding sites for

  10. Characterization of the starvation-induced chitinase CfcA and α-1,3-glucanase AgnB of Aspergillus niger

    NARCIS (Netherlands)

    van Munster, Jolanda M; Dobruchowska, Justyna M; Veloo, Ruud; Dijkhuizen, Lubbert; van der Maarel, Marc J E C

    2014-01-01

    The common saprophyte Aspergillus niger may experience carbon starvation in nature as well as during industrial fermentations. Starvation survival strategies, such as conidiation or the formation of exploratory hyphae, require energy and building blocks, which may be supplied by autolysis. Glycoside

  11. The role of the Aspergillus niger furin-type protease gene in processing of fungal proproteins and fusion proteins: Evidence for alternative processing of recombinant (fusion-) proteins

    NARCIS (Netherlands)

    Punt, P.J.; Drint-Kuijvenhoven, A.; Lokman, B.C.; Spencer, J.A.; Jeenes, D.; Archer, D.A.; Hondel, C.A.M.J.J. van den

    2003-01-01

    We have characterized growth and protein processing characteristics of Aspergillus niger strains carrying a disrupted allele of the previously cloned and characterized kexB gene [Appl. Environ. Microbiol. 66 (2000) 363] encoding a furin-type endoprotease. Deletion of the single-copy gene confirms it

  12. Comparative study of toxicity of azo dye Procion Red MX-5B following biosorption and biodegradation treatments with the fungi Aspergillus niger and Aspergillus terreus.

    Science.gov (United States)

    Almeida, E J R; Corso, C R

    2014-10-01

    Azo dyes are an important class of environmental contaminants and are characterized by the presence of one or more azo bonds (-N=N-) in their molecular structure. Effluents containing these compounds resist many types of treatments due to their molecular complexity. Therefore, alternative treatments, such as biosorption and biodegradation, have been widely studied to solve the problems caused by these substances, such as their harmful effects on the environment and organisms. The aim of the present study was to evaluate biosorption and biodegradation of the azo dye Procion Red MX-5B in solutions with the filamentous fungi Aspergillus niger and Aspergillus terreus. Decolorization tests were performed, followed by acute toxicity tests using Lactuca sativa seeds and Artemia salina larvae. Thirty percent dye removal of the solutions was achieved after 3 h of biosorption. UV-Vis spectroscopy revealed that removal of the dye molecules occurred without major molecular changes. The acute toxicity tests confirmed lack of molecular degradation following biosorption with A. niger, as toxicity to L. sativa seed reduced from 5% to 0%. For A. salina larvae, the solutions were nontoxic before and after treatment. In the biodegradation study with the fungus A. terreus, UV-Vis and FTIR spectroscopy revealed molecular degradation and the formation of secondary metabolites, such as primary and secondary amines. The biodegradation of the dye molecules was evaluated after 24, 240 and 336 h of treatment. The fungal biomass demonstrated considerable affinity for Procion Red MX-5B, achieving approximately 100% decolorization of the solutions by the end of treatment. However, the solutions resulting from this treatment exhibited a significant increase in toxicity, inhibiting the growth of L. sativa seeds by 43% and leading to a 100% mortality rate among the A. salina larvae. Based on the present findings, biodegradation was effective in the decolorization of the samples, but generated

  13. Differential Expression of Three α-Galactosidase Genes and a Single β-Galactosidase Gene from Aspergillus niger

    Science.gov (United States)

    de Vries, Ronald P.; van den Broeck, Hetty C.; Dekkers, Ester; Manzanares, Paloma; de Graaff, Leo H.; Visser, Jaap

    1999-01-01

    A gene encoding a third α-galactosidase (AglB) from Aspergillus niger has been cloned and sequenced. The gene consists of an open reading frame of 1,750 bp containing six introns. The gene encodes a protein of 443 amino acids which contains a eukaryotic signal sequence of 16 amino acids and seven putative N-glycosylation sites. The mature protein has a calculated molecular mass of 48,835 Da and a predicted pI of 4.6. An alignment of the AglB amino acid sequence with those of other α-galactosidases revealed that it belongs to a subfamily of α-galactosidases that also includes A. niger AglA. A. niger AglC belongs to a different subfamily that consists mainly of prokaryotic α-galactosidases. The expression of aglA, aglB, aglC, and lacA, the latter of which encodes an A. niger β-galactosidase, has been studied by using a number of monomeric, oligomeric, and polymeric compounds as growth substrates. Expression of aglA is only detected on galactose and galactose-containing oligomers and polymers. The aglB gene is expressed on all of the carbon sources tested, including glucose. Elevated expression was observed on xylan, which could be assigned to regulation via XlnR, the xylanolytic transcriptional activator. Expression of aglC was only observed on glucose, fructose, and combinations of glucose with xylose and galactose. High expression of lacA was detected on arabinose, xylose, xylan, and pectin. Similar to aglB, the expression on xylose and xylan can be assigned to regulation via XlnR. All four genes have distinct expression patterns which seem to mirror the natural substrates of the encoded proteins. PMID:10347026

  14. The weak acid preservative sorbic acid inhibits conidial germination and mycelial growth of Aspergillus niger through intracellular acidification.

    Science.gov (United States)

    Plumridge, Andrew; Hesse, Stephan J A; Watson, Adrian J; Lowe, Kenneth C; Stratford, Malcolm; Archer, David B

    2004-06-01

    The growth of the filamentous fungus Aspergillus niger, a common food spoilage organism, is inhibited by the weak acid preservative sorbic acid (trans-trans-2,4-hexadienoic acid). Conidia inoculated at 10(5)/ml of medium showed a sorbic acid MIC of 4.5 mM at pH 4.0, whereas the MIC for the amount of mycelia at 24 h developed from the same spore inoculum was threefold lower. The MIC for conidia and, to a lesser extent, mycelia was shown to be dependent on the inoculum size. A. niger is capable of degrading sorbic acid, and this ability has consequences for food preservation strategies. The mechanism of action of sorbic acid was investigated using (31)P nuclear magnetic resonance (NMR) spectroscopy. We show that a rapid decline in cytosolic pH (pH(cyt)) by more than 1 pH unit and a depression of vacuolar pH (pH(vac)) in A. niger occurs in the presence of sorbic acid. The pH gradient over the vacuole completely collapsed as a result of the decline in pH(cyt). NMR spectra also revealed that sorbic acid (3.0 mM at pH 4.0) caused intracellular ATP pools and levels of sugar-phosphomonoesters and -phosphodiesters of A. niger mycelia to decrease dramatically, and they did not recover. The disruption of pH homeostasis by sorbic acid at concentrations below the MIC could account for the delay in spore germination and retardation of the onset of subsequent mycelial growth.

  15. New insight into the disinfection mechanism of Fusarium monoliforme and Aspergillus niger by TiO2 photocatalyst under low intensity UVA light.

    Science.gov (United States)

    Pokhum, Chonlada; Viboonratanasri, Duangamon; Chawengkijwanich, Chamorn

    2017-11-01

    Titanium dioxide (TiO2) photocatalytic reaction has great potential for the disinfection of harmful pathogens. However, the disinfection mechanisms of TiO2 photocatalysis are not yet well-known for fungi and protozoa. In this work, the photocatalytic disinfection mechanism of Fusarium monoliforme and Aspergillus niger under low intensity UVA light (365nm, niger was more sensitive to UVA-light. Serious destructions of cell membrane and cellular organelles were shown in A. niger exposed to UVA-light only and photocatalytic treatments. However, morphological change in A. niger cell wall was only observed in photocatalytic treatment. Changes to the outermost melanin like layer and cell wall of A. niger spore due to photocatalytic treatment were greatly apparent while the intracellular organelles of A. niger spore were not affected. Therefore, regrowth of A. niger on agar plate was expected from the germination of A. niger spore in the subsequent dark. These observations give a better understanding of the photocatalytic disinfection mechanism toward fungi. Copyright © 2017 Elsevier B.V. All rights reserved.

  16. Characterization of a polyketide synthase in Aspergillus niger whose product is a precursor for both dihydroxynaphthalene (DHN) melanin and naphtho-γ-pyrone.

    Energy Technology Data Exchange (ETDEWEB)

    Chiang, Yi Ming; Meyer, Kristen M; Praseuth, Michael; Baker, Scott E; Bruno, Kenneth S; Wang, Clay C

    2010-12-06

    The genome sequencing of the fungus Aspergillus niger, an industrial workhorse, uncovered a large cache of genes encoding enzymes thought to be involved in the production of secondary metabolites yet to be identified. Identification and structural characterization of many of these predicted secondary metabolites are hampered by their low concentration relative to the known A. niger metabolites such as the naphtho-γ-pyrone family of polyketides. We deleted a nonreducing PKS gene in A. niger strain ATCC 11414, a daughter strain of A. niger ATCC strain 1015 whose genome was sequenced by the DOE Joint Genome Institute. This PKS encoding gene is a predicted ortholog of alb1 from Aspergillus fumigatus which is responsible for production of YWA1, a precursor of fungal DHN melanin. Our results show that the A. niger alb1 PKS is responsible for the production of the polyketide precursor for DHN melanin biosynthesis. Deletion of alb1 elimnates the production of major metabolites, naphtho-γ-pyrones. The generation of an A. niger strain devoid of naphtho-γ-pyrones will greatly facilitate the elucidation of cryptic biosynthetic pathways in this organism.

  17. Response surface optimization for enhanced production of cellulases with improved functional characteristics by newly isolated Aspergillus niger HN-2.

    Science.gov (United States)

    Oberoi, Harinder Singh; Rawat, Rekha; Chadha, Bhupinder Singh

    2014-01-01

    Fungi isolated from partially decayed wood log samples showing characteristic diversity for spore colour, colony morphology and arrangement of spores were assessed for cellulolytic enzyme production. Isolates showing a cellulolytic index of ≥2.0 were assayed for filter paper (FP) cellulase and β-glucosidase (BGL) production. Molecular characterization confirmed the identity of the selected cellulolytic isolate as a strain of Aspergillus niger (A. niger HN-2). Addition of 2 % (w/v) urea enhanced FP and BGL activity by about 20 and 60 %, respectively. Validation studies conducted at parameters (29 °C, pH 5.4, moisture content 72 % and 66 h) optimized through response surface methodology in a solid-state static tray fermentation resulted in FP, BGL, cellobiohydrolase I (CBHI), endoglucanase (EG), xylanase activity and protein content of 25.3 FPU/g ds, 750 IU/g ds, 13.2 IU/g ds, 190 IU/g ds, 2890 IU/g ds and 0.9 mg/ml, respectively. In comparison, A. niger N402 which is a model organism for growth and development studies, produced significantly lower FP, BGL, CBHI, EG, xylanase activity and protein content of 10.0 FPU/g ds, 100 IU/g ds, 2.3 IU/g ds, 50 IU/g ds, 500 IU/g ds and 0.75 mg/ml, respectively under the same process conditions as were used for A. niger HN-2. Process optimization led to nearly 1.8- and 2.2-fold increase in FP and BGL activity, respectively showing promise for cellulase production by A. niger HN-2 at a higher scale of operation. Zymogram analysis revealed two isoforms each for EG and cellobiohydrolase and three isoforms for BGL. Crude cellulase complex produced by A. niger HN-2 exhibited thermostability under acidic conditions showing potential for use in biofuel industry.

  18. Production of plant cell wall degrading enzymes by monoculture and co-culture of Aspergillus niger and Aspergillus terreus under SSF of banana peels.

    Science.gov (United States)

    Rehman, Shazia; Aslam, Hina; Ahmad, Aqeel; Khan, Shakeel Ahmed; Sohail, Muhammad

    2014-01-01

    Filamentous fungi are considered to be the most important group of microorganisms for the production of plant cell wall degrading enzymes (CWDE), in solid state fermentations. In this study, two fungal strains Aspergillus niger MS23 and Aspergillus terreus MS105 were screened for plant CWDE such as amylase, pectinase, xylanase and cellulases (β-glucosidase, endoglucanase and filterpaperase) using a novel substrate, Banana Peels (BP) for SSF process. This is the first study, to the best of our knowledge, to use BP as SSF substrate for plant CWDE production by co-culture of fungal strains. The titers of pectinase were significantly improved in co-culture compared to mono-culture. Furthermore, the enzyme preparations obtained from monoculture and co-culture were used to study the hydrolysis of BP along with some crude and purified substrates. It was observed that the enzymatic hydrolysis of different crude and purified substrates accomplished after 26 h of incubation, where pectin was maximally hydrolyzed by the enzyme preparations of mono and co-culture. Along with purified substrates, crude materials were also proved to be efficiently degraded by the cocktail of the CWDE. These results demonstrated that banana peels may be a potential substrate in solid-state fermentation for the production of plant cell wall degrading enzymes to be used for improving various biotechnological and industrial processes.

  19. Production of plant cell wall degrading enzymes by monoculture and co-culture of Aspergillus niger and Aspergillus terreus under SSF of banana peels

    Directory of Open Access Journals (Sweden)

    Shazia Rehman

    2014-12-01

    Full Text Available Filamentous fungi are considered to be the most important group of microorganisms for the production of plant cell wall degrading enzymes (CWDE, in solid state fermentations. In this study, two fungal strains Aspergillus niger MS23 and Aspergillus terreus MS105 were screened for plant CWDE such as amylase, pectinase, xylanase and cellulases (β-glucosidase, endoglucanase and filterpaperase using a novel substrate, Banana Peels (BP for SSF process. This is the first study, to the best of our knowledge, to use BP as SSF substrate for plant CWDE production by co-culture of fungal strains. The titers of pectinase were significantly improved in co-culture compared to mono-culture. Furthermore, the enzyme preparations obtained from monoculture and co-culture were used to study the hydrolysis of BP along with some crude and purified substrates. It was observed that the enzymatic hydrolysis of different crude and purified substrates accomplished after 26 h of incubation, where pectin was maximally hydrolyzed by the enzyme preparations of mono and co-culture. Along with purified substrates, crude materials were also proved to be efficiently degraded by the cocktail of the CWDE. These results demonstrated that banana peels may be a potential substrate in solid-state fermentation for the production of plant cell wall degrading enzymes to be used for improving various biotechnological and industrial processes.

  20. Produção de celulases por Aspergillus niger e cinética da desativação celulásica=Cellulases production by Aspergillus niger and cellulase deactivation kinetic

    Directory of Open Access Journals (Sweden)

    Caroline Mariana de Aguiar

    2011-10-01

    Full Text Available O presente trabalho teve como objetivo a avaliação da cinética de produção de enzimas celulases pelo fungo Aspergillus niger e da cinética de desativação das celulases. Foi utilizado bagaço de cana-de-açúcar pré-tratado como fonte de carbono na fermentação para a produção do complexo celulásico e também como substrato da hidrólise enzimática. A. niger foi cultivado em três bateladas, cada uma contendo 10, 50 e 100 g L-1 de bagaço pré-tratado com NaOH 4% (m v-1. A cinética da produção das celulases foi obtida determinando-se a atividade enzimática das amostras coletadas ao longo do tempo. As variações do pH também foram determinadas. A deativação enzimática foi avaliada determinando-se periodicamente a atividade das amostras armazenadas nas condições de resfriamento (4°C e de congelamento (-18ºC. Conclui-se que o A. niger produz celulases quando cultivado em meio de cultivo contendo bagaço de cana-de-açúcar pré-tratado e que o tempo ideal para coleta do caldo enzimático foi de aproximadamente sete dias, com produtividade máxima de 0,0013 U mL-1∙h para a batelada com 10 g L-1 e 0,0018 U mL-1∙h para as bateladas com 50 e 100 g L-1. O complexo celulásico não sofre desativação se armazenado à temperatura de -18°C por 43 dias, mas perde cerca de 40% da sua atividade após 48h se armazenado a 4°C.This work aimed to evaluate the kinetic for the cellulase production by Aspergillus niger and the deactivation kinetic of the cellulase enzymes. Cellulase were produced in three different batches using NaOH 4% (w v-1 pre-treated sugarcane bagasse as the carbon source in the fermentation broth. The amount of the bagasse in each batch was 10, 50 and 100 g L-1. The kinetic of the cellulase production was accomplished by periodically determining the cellulasic activity of the fermentation broth using pre-treated bagasse as the hydrolysis substrate. Changes in the pH also were determined. The cellulase

  1. Treatment of APMP pulping effluent based on aerobic fermentation with Aspergillus niger and post-coagulation/flocculation.

    Science.gov (United States)

    Liu, Tingzhi; He, Zhibin; Hu, Huiren; Ni, Yonghao

    2011-04-01

    A novel two-stage biological/flocculation process was developed for treating the pulping effluent from the alkaline peroxide mechanical pulping (APMP) process. In the first biological stage, the aerobic fermentation by using Aspergillus niger can decrease the chemical oxygen demand (COD) by about 60% while producing about 7 g/l of solid biomass. In the second stage (post-coagulation/flocculation), the residual COD, turbidity and color, can be further decreased by using alum and polyacrylamide (PAM). The overall removal efficiencies of COD, color and turbidity from the APMP pulping effluent by the above two-stage biological-coagulation/flocculation process were 93%, 92% and 99%, respectively, under the conditions studied. Copyright © 2011 Elsevier Ltd. All rights reserved.

  2. Caracterización de la biomasa inactiva de Aspergillus niger O-5 como sorbente de Pb (II

    Directory of Open Access Journals (Sweden)

    Yusleydi Enamorado Horrutiner

    2011-01-01

    Full Text Available The inactive biomass of fungus Aspergillus niger O-5 obtained in Cuba was characterized as sorbent of Pb2+ by several structural analysis and others techniques. In addition, the biomass was studied for the separation / preconcentration of Pb2+ from aqueous solution. The maximum biosorption capacity was obtained for the contact time of 30 min and pH 5. The kinetic of sorption process occurred according to the model of Ho. The Freundlich or Langmuir models suitably described the experimental adsorption isotherms. The biomass can be used as sorbent for Pb2+ with a maximum capacity of 4.7 - 6.2 mg g-1. The pretreatment with NaOH solution improved its sorption capacity.

  3. Treatment of African catfish, Clarias gariepinus wastewater utilizing phytoremediation of microalgae, Chlorella sp. with Aspergillus niger bio-harvesting.

    Science.gov (United States)

    Nasir, Nurfarahana Mohd; Bakar, Nur Syuhada Abu; Lananan, Fathurrahman; Abdul Hamid, Siti Hajar; Lam, Su Shiung; Jusoh, Ahmad

    2015-08-01

    This study focuses on the evaluation of the performance of Chlorella sp. in removing nutrient in aquaculture wastewater and its correlation with the kinetic growth of Chlorella sp. The treatment was applied with various Chlorella sp. inoculation dosage ranging from 0% to 60% (v/v) of wastewater. The optimum inoculation dosage was recorded at 30% (v/v) with effluent concentration of ammonia and orthophosphate recording at 0.012mgL(-1) and 0.647mgL(-1), respectively on Day 11. The optimum dosage for bio-flocculation process was obtained at 30mgL(-1) of Aspergillus niger with a harvesting efficiency of 97%. This type of development of phytoremediation with continuous bio-harvesting could promote the use of sustainable green technology for effective wastewater treatment. Copyright © 2015 Elsevier Ltd. All rights reserved.

  4. Influência da glicose sobre o consumo de fenol por Aspergillus niger an 400 em reatores em batelada

    OpenAIRE

    Rodrigues,Kelly de Araújo; Sampaio,Glória Maria Marinho S.; Zaiat,Marcelo; Santaella,Sandra Tédde

    2007-01-01

    Aspergillus niger AN 400 foi inoculado em reatores em batelada para remoção de fenol de meio sintético, na presença e ausência de glicose. O experimento possuía: 5 reatores de controle (grupo 1) com meio contento apenas fenol; 5 reatores (grupo 2) inoculados com fungos e com meio com fenol e sem glicose; e 5 reatores (grupo 3) inoculados com fungos e com meio com fenol e glicose (5 g/L). Os reatores foram agitados a 200 rpm, a 30°C, durante 5 dias. A concentração média inicial de fenol foi de...

  5. INFLUENCE OF SOLID MOISTURE AND BED HEIGHT ON CULTIVATION OF Aspergillus niger FROM SUGARCANE BAGASSE WITH VINASSE

    Directory of Open Access Journals (Sweden)

    R. G. Bastos

    2015-06-01

    Full Text Available AbstractSolid-state cultivation (SSC may be defined as growth of microorganisms on a solid support impregnated or not with a nutrient solution in near absence of free-water conditions. The use of sugarcane bagasse as a support for SSC usually demands that the particles are impregnated and moistened with nutrient solution. Vinasse is the main wastewater of ethanol fermentation-distillation. As there are no reports of the use of wastewater for moistening solid supports in SSC, the proposal is the development of an innovative process, with valuation of these by-products. Thus, the aim of this research was to evaluate SSC of Aspergillus niger using sugarcane bagasse and vinasse for citric acid production. The results indicate that citric acid production and glucose consumption are dependent on oxygen availability, which can be modulated by selection of bed height and air-flow in packed-bed bioreactors.

  6. Acid protease and formation of multiple forms of glucoamylase in batch and continuous cultures of Aspergillus niger

    DEFF Research Database (Denmark)

    Aalbæk, Thomas; Reeslev, Morten; Jensen, Bo

    2002-01-01

    In order to identify factors responsible for production of multiple forms of glucoamylase (GA) by Aspergillus niger Bo-1, the fungus was cultured in both complex and defined media in pH-controlled batch fermenters and chemostats. At all culture conditions three forms of GA were produced...... was cultivated in batch and continuous fermentations with a constant pH of 3.0 or 5.0, whereas acidification of both complex and defined batch culture media after 20 h of growth induced a significant secretion of acid protease(s). The protease(s) present in the complex medium catalysed modification...... with molecular weights of approx. 91 (GAI), 73 (GAII), and 59 kDa (GAIII). Data from batch fermentations with constant pH 3.0 and 5.0 showed a uniform distribution of extracellular GA forms throughout the fermentations and independent of culture growth phases. Furthermore, steady-state data from chemostat...

  7. Acid protease and formation of multiple forms of glycoamylase in batch and continuous cultures of Aspergillus niger

    DEFF Research Database (Denmark)

    Aalbæk, Thomas; Reeslev, Morten; Jensen, Bo

    2002-01-01

    In order to identify factors responsible for production of multiple forms of glucoamylase (GA) by Aspergillus niger Bo-1, the fungus was cultured in both complex and defined media in pH-controlled batch fermenters and chemostats. At all culture conditions three forms of GA were produced...... was cultivated in batch and continuous fermentations with a constant pH of 3.0 or 5.0, whereas acidification of both complex and defined batch culture media after 20 h of growth induced a significant secretion of acid protease(s). The protease(s) present in the complex medium catalysed modification...... with molecular weights of approx. 91 (GAI), 73 (GAII), and 59 kDa (GAIII). Data from batch fermentations with constant pH 3.0 and 5.0 showed a uniform distribution of extracellular GA forms throughout the fermentations and independent of culture growth phases. Furthermore, steady-state data from chemostat...

  8. High titer gluconic acid fermentation by Aspergillus niger from dry dilute acid pretreated corn stover without detoxification.

    Science.gov (United States)

    Zhang, Hongsen; Zhang, Jian; Bao, Jie

    2016-03-01

    This study reported a high titer gluconic acid fermentation using dry dilute acid pretreated corn stover (DDAP) hydrolysate without detoxification. The selected fermenting strain Aspergillus niger SIIM M276 was capable of inhibitor degradation thus no detoxification on pretreated corn stover was required. Parameters of gluconic acid fermentation in corn stover hydrolysate were optimized in flasks and in fermentors to achieve 76.67 g/L gluconic acid with overall yield of 94.91%. The sodium gluconate obtained from corn stover was used as additive for extending setting time of cement mortar and similar function was obtained with starch based sodium gluconate. This study provided the first high titer gluconic acid production from lignocellulosic feedstock with potential of industrial applications. Copyright © 2015 Elsevier Ltd. All rights reserved.

  9. [Cloning of feruloyl esterase gene from Aspergillus niger h408 and high-efficient expression in Pichia pastoris].

    Science.gov (United States)

    Zhou, Yanyan; Liu, Xinli; Chen, Jing; Hu, Hongyu; Hou, Yunhua

    2014-08-04

    To achieve the high-efficiency expression of feruloyl estrase gene (AnfaeA) from Aspergillus niger h408 in Pichia pastoris and characterize the recombinant feruloyl esterase (FAE). Using gene splicing by overlap extension (SOE), we cloned AnfaeA gene from A. niger h408 and subcloned into T vector for sequencing analysis. The expression vector pPIC9K-Anfae was constructed by the ligation of the Anfae A gene into the shuttle vector pPIC9K. The plasmid pPIC9K-Anfae was linearized and then electrotransformed into P. pastoris GS115. The recombinant strain with high level of FAE activity was obtained through plate screening. Effects of pH and temperature on recombinant FAE were determined by ultraviolet (UV) methods. We have successfully cloned and high-efficiently expressed the AnfaeA gene (GenBank: KF911349) from A. niger h408 in P. pastoris GS115. The sequencing result showed that the length of Anfae A was 783bp. The gene contained an Open Reading Frame encoding 260 amino acids and was similar to feruloyl esterase A from A. niger by homology analysis. The deduced amino acids contained a typical active lid and catalytic triad of lipase. The SDS-PAGE result indicated that molecular weight of the recombinant FAE was about 30 kDa and the activity of the recombinant enzyme was 24.72 U/mL. The specific activity of the recombinant FAE was 40.84 U/mg. Compared with A. niger h408, the recombinant enzyme activity increased about to 1100 times. The optimal temperature and pH for recombinant FAE was 50 degrees C and 5.0, respectively. Recombinant FAE showed nearly 80% of its maximal activity at 60 degrees C and was active in the pH range 4.0-9.0. The high-efficient expression of AnfaeA gene in P. pastoris provided a prerequisite for achieving industrial application in feed and paper-making industry. In addition, the results established the experimental basis for further improvement of recombinant feruloyl esterase by directed evolution.

  10. Comparative genomics of citric-acid producing Aspergillus niger ATCC 1015 versus enzyme-producing CBS 513.88

    Energy Technology Data Exchange (ETDEWEB)

    Grigoriev, Igor V.; Baker, Scott E.; Andersen, Mikael R.; Salazar, Margarita P.; Schaap, Peter J.; Vondervoot, Peter J.I. van de; Culley, David; Thykaer, Jette; Frisvad, Jens C.; Nielsen, Kristen F.; Albang, Richard; Albermann, Kaj; Berka, Randy M.; Braus, Gerhard H.; Braus-Stromeyer, Susanna A.; Corrochano, Luis M.; Dai, Ziyu; Dijck, Piet W.M. van; Hofmann, Gerald; Lasure, Linda L.; Magnusson, Jon K.; Meijer, Susan L.; Nielsen, Jakob B.; Nielsen, Michael L.; Ooyen, Albert J.J. van; Panther, Kathyrn S.; Pel, Herman J.; Poulsen, Lars; Samson, Rob A.; Stam, Hen; Tsang, Adrian; Brink, Johannes M. van den; Atkins, Alex; Aerts, Andrea; Shapiro, Harris; Pangilinan, Jasmyn; Salamov, Asaf; Lou, Yigong; Lindquist, Erika; Lucas, Susan; Grimwood, Jane; Kubicek, Christian P.; Martinez, Diego; Peij, Noel N.M.E. van; Roubos, Johannes A.; Nielsen, Jens

    2011-04-28

    The filamentous fungus Aspergillus niger exhibits great diversity in its phenotype. It is found globally, both as marine and terrestrial strains, produces both organic acids and hydrolytic enzymes in high amounts, and some isolates exhibit pathogenicity. Although the genome of an industrial enzyme-producing A. niger strain (CBS 513.88) has already been sequenced, the versatility and diversity of this species compels additional exploration. We therefore undertook whole genome sequencing of the acidogenic A. niger wild type strain (ATCC 1015), and produced a genome sequence of very high quality. Only 15 gaps are present in the sequence and half the telomeric regions have been elucidated. Moreover, sequence information from ATCC 1015 was utilized to improve the genome sequence of CBS 513.88. Chromosome-level comparisons uncovered several genome rearrangements, deletions, a clear case of strain-specific horizontal gene transfer, and identification of 0.8 megabase of novel sequence. Single nucleotide polymorphisms per kilobase (SNPs/kb) between the two strains were found to be exceptionally high (average: 7.8, maximum: 160 SNPs/kb). High variation within the species was confirmed with exo-metabolite profiling and phylogenetics. Detailed lists of alleles were generated, and genotypic differences were observed to accumulate in metabolic pathways essential to acid production and protein synthesis. A transcriptome analysis revealed up-regulation of the electron transport chain, specifically the alternative oxidative pathway in ATCC 1015, while CBS 513.88 showed significant up-regulation of genes relevant to glucoamylase A production, such as tRNA-synthases and protein transporters. Our results and datasets from this integrative systems biology analysis resulted in a snapshot of fungal evolution and will support further optimization of cell factories based on filamentous fungi.[Supplemental materials (10 figures, three text documents and 16 tables) have been made available

  11. Involvement of the opportunistic pathogen Aspergillus tubingensis in osteomyelitis of the maxillary bone: A case report

    NARCIS (Netherlands)

    Bathoorn, E.; Escobar Salazar, N.; Sepehrkhouy, S.; Meijer, M.; de Cock, H.; Haas, P.J.

    2013-01-01

    Background: Aspergillus tubingensis is a black Aspergillus belonging to the Aspergillus section Nigri, which includes species that morphologically resemble Aspergillus niger. Recent developments in species determination have resulted in clinical isolates presumed to be Aspergillus niger being

  12. Cloning, expression, purification, crystallization and preliminary X-ray diffraction analysis of variants of monoamine oxidase from Aspergillus niger

    Energy Technology Data Exchange (ETDEWEB)

    Atkin, Kate E. [Structural Biology Laboratory, Department of Chemistry, University of York, York YO10 5YW (United Kingdom); Reiss, Renate; Turner, Nicholas J. [School of Chemistry, Manchester Interdisciplinary Biocentre, University of Manchester, 131 Princess Street, Manchester M1 7DN (United Kingdom); Brzozowski, Andrzej M.; Grogan, Gideon, E-mail: grogan@ysbl.york.ac.uk [Structural Biology Laboratory, Department of Chemistry, University of York, York YO10 5YW (United Kingdom)

    2008-03-01

    Crystals of A. niger monoamine oxidase variants display P2{sub 1} or P4{sub 1}2{sub 1}2/P4{sub 3}2{sub 1}2 symmetry, with eight or two molecules in the asymmetric unit, respectively. Monoamine oxidase from Aspergillus niger (MAO-N) is an FAD-dependent enzyme that catalyses the conversion of terminal amines to their corresponding aldehydes. Variants of MAO-N produced by directed evolution have been shown to possess altered substrate specificity. Crystals of two of these variants (MAO-N-3 and MAO-N-5) have been obtained; the former displays P2{sub 1} symmetry with eight molecules per asymmetric unit and the latter has P4{sub 1}2{sub 1}2 or P4{sub 3}2{sub 1}2 symmetry and two molecules per asymmetric unit. Solution of these structures will help shed light on the molecular determinants of improved activity and high enantioselectivity towards a broad range of substrates.

  13. Optimization of the parameters for decolourization by Aspergillus niger of anaerobically digested distillery spentwash pretreated with polyaluminium chloride.

    Science.gov (United States)

    Singh, S S; Dikshit, A K

    2010-04-15

    Molasses spentwash from distilleries is characterized by high COD and colour. The fungal decolourization of anaerobically digested molasses spentwash requires significant dilution. In this study, decolourization by Aspergillus niger isolate IITB-V8 was performed on polyaluminium chloride (PAC) treated anaerobically digested spentwash without dilution of wastewater. Optimization of parameters was studied using statistical experimental designs. In the first step, Plackett-Burman design was used for screening the important parameters. Glucose was taken as the carbon source for the growth of A. niger. KH(2)PO(4) and pH were found to be the important factors affecting decolourization. In the second step, Box-Behnken design was used to determine the optimum level of each of the significant parameters. A second-order polynomial was determined by the multiple regression analysis of the experimental data. The optimum values for the important factors to achieve maximum decolourization of 68.4% were 5.5 g/L Glucose, 1.2 g/L KH(2)PO(4) and 5 pH. The determination coefficient (R(2)) was 0.9973, which ensures adequate credibility of the model. The total decolourization obtained after fungal treatment was 86.8% which indicates fungal decolourization after pretreatment with PAC is a viable option for the treatment of digested molasses spentwash. 2009 Elsevier B.V. All rights reserved.

  14. Lipase Production in Solid-State Fermentation Monitoring Biomass Growth of Aspergillus niger Using Digital Image Processing

    Science.gov (United States)

    Dutra, Julio C. V.; da Terzi, Selma C.; Bevilaqua, Juliana Vaz; Damaso, Mônica C. T.; Couri, Sônia; Langone, Marta A. P.; Senna, Lilian F.

    The aim of this study was to monitor the biomass growth of Aspergillus niger in solid-state fermentation (SSF) for lipase production using digital image processing technique. The strain A. niger 11T53A14 was cultivated in SSF using wheat bran as support, which was enriched with 0.91% (m/v) of ammonium sulfate. The addition of several vegetable oils (castor, soybean, olive, corn, and palm oils) was investigated to enhance lipase production. The maximum lipase activity was obtained using 2% (m/m) castor oil. In these conditions, the growth was evaluated each 24 h for 5 days by the glycosamine content analysis and digital image processing. Lipase activity was also determined. The results indicated that the digital image process technique can be used to monitor biomass growth in a SSF process and to correlate biomass growth and enzyme activity. In addition, the immobilized esterification lipase activity was determined for the butyl oleate synthesis, with and without 50% v/v hexane, resulting in 650 and 120 U/g, respectively. The enzyme was also used for transesterification of soybean oil and ethanol with maximum yield of 2.4%, after 30 min of reaction.

  15. Role of Aspergillus niger acrA in Arsenic Resistance and Its Use as the Basis for an Arsenic Biosensor

    Science.gov (United States)

    Choe, Se-In; Gravelat, Fabrice N.; Al Abdallah, Qusai; Lee, Mark J.; Gibbs, Bernard F.

    2012-01-01

    Arsenic contamination of groundwater sources is a major issue worldwide, since exposure to high levels of arsenic has been linked to a variety of health problems. Effective methods of detection are thus greatly needed as preventive measures. In an effort to develop a fungal biosensor for arsenic, we first identified seven putative arsenic metabolism and transport genes in Aspergillus niger, a widely used industrial organism that is generally regarded as safe (GRAS). Among the genes tested for RNA expression in response to arsenate, acrA, encoding a putative plasma membrane arsenite efflux pump, displayed an over 200-fold increase in gene expression in response to arsenate. We characterized the function of this A. niger protein in arsenic efflux by gene knockout and confirmed that AcrA was located at the cell membrane using an enhanced green fluorescent protein (eGFP) fusion construct. Based on our observations, we developed a putative biosensor strain containing a construct of the native promoter of acrA fused with egfp. We analyzed the fluorescence of this biosensor strain in the presence of arsenic using confocal microscopy and spectrofluorimetry. The biosensor strain reliably detected both arsenite and arsenate in the range of 1.8 to 180 μg/liter, which encompasses the threshold concentrations for drinking water set by the World Health Organization (10 and 50 μg/liter). PMID:22467499

  16. Efficient Expression of an Acidic Endo-polygalacturonase from Aspergillus niger and Its Application in Juice Production.

    Science.gov (United States)

    Wang, Jiaojiao; Zhang, Yuhong; Qin, Xing; Gao, Lingyu; Han, Bin; Zhang, Deqing; Li, Jinyang; Huang, He; Zhang, Wei

    2017-04-05

    An endo-polygalacturonase gene (pga-zj5a) was cloned by reverse transcription from cDNAs synthesized from Aspergillus niger ZJ5 total RNA. The open reading frame of pga-zj5a was 1089 base pairs encoding 362 amino acids. Pga-zj5a lacking a signal peptide sequence was successfully amplified using A. niger ZJ5 cDNA as the template and was ligated into the pPIC9 vector. The resulting plasmid was transformed into competent cells of Pichia pastoris GS115 for heterologous expression. The polygalacturonase showed a maximum activity level of 10436 U/mL in the culture supernatant from a 3 L fermenter. Assays of enzymatic properties showed that the optimal pH and temperature of the recombinant PGA-ZJ5A were 4.5 and 40 °C, respectively. PGA-ZJ5A was effective in pear juice clarification, increased the volume of pear juice by 41.8%, and improved its light transmittance 3-fold.

  17. Expanding the feruloyl esterase gene family of Aspergillus niger by characterization of a feruloyl esterase, FaeC.

    Science.gov (United States)

    Dilokpimol, Adiphol; Mäkelä, Miia R; Mansouri, Sadegh; Belova, Olga; Waterstraat, Martin; Bunzel, Mirko; de Vries, Ronald P; Hildén, Kristiina S

    2017-07-25

    A feruloyl esterase (FAE) from Aspergillus niger N402, FaeC was heterologously produced in Pichia pastoris X-33 in a yield of 10mg/L. FaeC was most active at pH 7.0 and 50°C, and showed broad substrate specificity and catalyzed the hydrolysis of methyl 3,4-dimethoxycinnamate, ethyl ferulate, methyl ferulate, methyl p-coumarate, ethyl coumarate, methyl sinapate, and methyl caffeate. The enzyme released both ferulic acid and p-coumaric acid from wheat arabinoxylan and sugar beet pectin (up to 3mg/g polysaccharide), and acted synergistically with a commercial xylanase increasing the release of ferulic acid up to six-fold. The expression of faeC increased over time in the presence of feruloylated polysaccharides. Cinnamic, syringic, caffeic, vanillic and ferulic acid induced the expression of faeC. Overall expression of faeC was very low in all tested conditions, compared to two other A. niger FAE encoding genes, faeA and faeB. Our data showed that the fae genes responded differently towards the feruloylated polysaccharides and tested monomeric phenolic compounds suggesting that the corresponding FAE isoenzymes may target different substrates in a complementary manner. This may increase the efficiency of the degradation of diverse plant biomass. Copyright © 2017 Elsevier B.V. All rights reserved.

  18. Isolation, molecular cloning and expression of cellobiohydrolase B (CbhB) from Aspergillus niger in Escherichia coli

    Science.gov (United States)

    Woon, J. S. K.; Murad, A. M. A.; Abu Bakar, F. D.

    2015-09-01

    A cellobiohydrolase B (CbhB) from Aspergillus niger ATCC 10574 was cloned and expressed in E. coli. CbhB has an open reading frame of 1611 bp encoding a putative polypeptide of 536 amino acids. Analysis of the encoded polypeptide predicted a molecular mass of 56.2 kDa, a cellulose binding module (CBM) and a catalytic module. In order to obtain the mRNA of cbhB, total RNA was extracted from A. niger cells induced by 1% Avicel. First strand cDNA was synthesized from total RNA via reverse transcription. The full length cDNA of cbhB was amplified by PCR and cloned into the cloning vector, pGEM-T Easy. A comparison between genomic DNA and cDNA sequences of cbhB revealed that the gene is intronless. Upon the removal of the signal peptide, the cDNA of cbhB was cloned into the expression vector pET-32b. However, the recombinant CbhB was expressed in Escherichia coli Origami DE3 as an insoluble protein. A homology model of CbhB predicted the presence of nine disulfide bonds in the protein structure which may have contributed to the improper folding of the protein and thus, resulting in inclusion bodies in E. coli.

  19. KUALITAS PROTEIN DAN KOMPOSISI ASAM AMINO AMPAS SAGU HASIL FERMENTASI ASPERGILLUS NIGER DENGAN PENAMBAHAN UREA DAN ZEOLIT

    Directory of Open Access Journals (Sweden)

    La Ode Muhsafaat

    2015-08-01

    Full Text Available This experiment was designed to evaluate protein quality and amino acid composition of fermented sago pulp by Aspergillus niger with urea and zeolit addition at different levels. The experiment was design in factorial completely randomized with two factors and three replicates. The first factor was levels of urea addition (0, 2.5, and 5%, the second factor was levels of zeolite addition (0, 2.5, and 5% and used 2% A. niger on sago pulp (dry matter form. Variables observed were the levels of crude protein and amino acids. Data were tested using descriptive and Analysis of Variance (ANOVA, the differences among treatments means examined by Duncan Multiple Range Test. The results howed that there was interaction between levels of addition urea and zeolite on crude protein value. The addition urea and zeolite at 5% had the high increased 15.49 ± 0.33% to crude protein content. The amino acid content of fermented sago pulp descriptively increased in urea treatment, but didn’t show increased in zeolite treatment. Based on the calculation of the score of chemical and index essential amino acids, fermented sago pulp at 5% urea treatment has a chemical score of 17.86, 18.01, and 11.85 with the essential amino acid deficient is leucine, and has an index of 38.33, 36.43, and 28.45%.

  20. Isolation, molecular cloning and expression of cellobiohydrolase B (CbhB) from Aspergillus niger in Escherichia coli

    Energy Technology Data Exchange (ETDEWEB)

    Woon, J. S. K., E-mail: jameswoon@siswa.ukm.edu.my; Murad, A. M. A., E-mail: munir@ukm.edu.my; Abu Bakar, F. D., E-mail: fabyff@ukm.edu.my [School of Biosciences and Biotechnology, Faculty of Science and Technology, Universiti Kebangsaan Malaysia, 43600 UKM Bangi, Selangor (Malaysia)

    2015-09-25

    A cellobiohydrolase B (CbhB) from Aspergillus niger ATCC 10574 was cloned and expressed in E. coli. CbhB has an open reading frame of 1611 bp encoding a putative polypeptide of 536 amino acids. Analysis of the encoded polypeptide predicted a molecular mass of 56.2 kDa, a cellulose binding module (CBM) and a catalytic module. In order to obtain the mRNA of cbhB, total RNA was extracted from A. niger cells induced by 1% Avicel. First strand cDNA was synthesized from total RNA via reverse transcription. The full length cDNA of cbhB was amplified by PCR and cloned into the cloning vector, pGEM-T Easy. A comparison between genomic DNA and cDNA sequences of cbhB revealed that the gene is intronless. Upon the removal of the signal peptide, the cDNA of cbhB was cloned into the expression vector pET-32b. However, the recombinant CbhB was expressed in Escherichia coli Origami DE3 as an insoluble protein. A homology model of CbhB predicted the presence of nine disulfide bonds in the protein structure which may have contributed to the improper folding of the protein and thus, resulting in inclusion bodies in E. coli.